CA2656648A1 - Self-assembled, micropatterned, and radio frequency (rf) shielded biocontainers and their uses for remote spatially controlled chemical delivery - Google Patents
Self-assembled, micropatterned, and radio frequency (rf) shielded biocontainers and their uses for remote spatially controlled chemical delivery Download PDFInfo
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- CA2656648A1 CA2656648A1 CA002656648A CA2656648A CA2656648A1 CA 2656648 A1 CA2656648 A1 CA 2656648A1 CA 002656648 A CA002656648 A CA 002656648A CA 2656648 A CA2656648 A CA 2656648A CA 2656648 A1 CA2656648 A1 CA 2656648A1
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- 229910052618 mica group Inorganic materials 0.000 description 1
- 238000004377 microelectronic Methods 0.000 description 1
- 239000004005 microsphere Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229910052750 molybdenum Inorganic materials 0.000 description 1
- 239000011733 molybdenum Substances 0.000 description 1
- 238000000465 moulding Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- CXQXSVUQTKDNFP-UHFFFAOYSA-N octamethyltrisiloxane Chemical compound C[Si](C)(C)O[Si](C)(C)O[Si](C)(C)C CXQXSVUQTKDNFP-UHFFFAOYSA-N 0.000 description 1
- 229940099990 ogen Drugs 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 238000000879 optical micrograph Methods 0.000 description 1
- 230000003534 oscillatory effect Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052763 palladium Inorganic materials 0.000 description 1
- 230000005298 paramagnetic effect Effects 0.000 description 1
- 230000010412 perfusion Effects 0.000 description 1
- KJFMBFZCATUALV-UHFFFAOYSA-N phenolphthalein Chemical compound C1=CC(O)=CC=C1C1(C=2C=CC(O)=CC=2)C2=CC=CC=C2C(=O)O1 KJFMBFZCATUALV-UHFFFAOYSA-N 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 238000004987 plasma desorption mass spectroscopy Methods 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 229920001993 poloxamer 188 Polymers 0.000 description 1
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 description 1
- 229920000573 polyethylene Polymers 0.000 description 1
- 229920001296 polysiloxane Polymers 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- ZMRUPTIKESYGQW-UHFFFAOYSA-N propranolol hydrochloride Chemical compound [H+].[Cl-].C1=CC=C2C(OCC(O)CNC(C)C)=CC=CC2=C1 ZMRUPTIKESYGQW-UHFFFAOYSA-N 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000013102 re-test Methods 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 239000012429 reaction media Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000001172 regenerating effect Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 231100000241 scar Toxicity 0.000 description 1
- 238000005204 segregation Methods 0.000 description 1
- 230000001235 sensitizing effect Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 108010080511 serum sodium transport inhibitor Proteins 0.000 description 1
- 239000008279 sol Substances 0.000 description 1
- 239000011343 solid material Substances 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000002344 surface layer Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 229940056345 tums Drugs 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
- A61K9/0024—Solid, semi-solid or solidifying implants, which are implanted or injected in body tissue
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0087—Galenical forms not covered by A61K9/02 - A61K9/7023
- A61K9/0097—Micromachined devices; Microelectromechanical systems [MEMS]; Devices obtained by lithographic treatment of silicon; Devices comprising chips
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y30/00—Nanotechnology for materials or surface science, e.g. nanocomposites
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Nanotechnology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Diabetes (AREA)
- Animal Behavior & Ethology (AREA)
- Dermatology (AREA)
- Crystallography & Structural Chemistry (AREA)
- Epidemiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Chemical & Material Sciences (AREA)
- General Physics & Mathematics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biomedical Technology (AREA)
- Neurosurgery (AREA)
- Obesity (AREA)
- Hematology (AREA)
- Physics & Mathematics (AREA)
- Composite Materials (AREA)
- Condensed Matter Physics & Semiconductors (AREA)
- Organic Chemistry (AREA)
- Materials Engineering (AREA)
- Endocrinology (AREA)
- Emergency Medicine (AREA)
- Biophysics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Medical Informatics (AREA)
- Molecular Biology (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Medicinal Preparation (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The present invention relates to a nanoscale or microscale particle for encapsulation and delivery of materials or substances, including, but not limited to, cells, drugs, tissue, gels and polymers contained within the particle, with subsequent release of the therapeutic materials in situ, methods of fabricating the particle by folding a 2D precursor into the 3D particle, and the use of the particle in in-vivo or in-vitro applications The particle can be in any polyhedral shape and its surfaces can have either no perforations or nano/microscale perforations The particle is coated with a biocompatible metal, e g gold, or polymer. e g parvlene, layer and the surfaces and hinges of the particle are made of any metal or polymer combinations.
Description
-Se1f Assembted. Wcr0 atterned 4inti Radi0 Frec u~nc> R1F+` Shieldeci 113-raC0vitai~~ers atuc.ll `C`heirlses t`or :Remote Saat.iallj, Controlled C:heiiii+eat Deliverk CROSS-REFERENCE TO RELATED APPLICATIONS
100011 Tl7B application claims the ber~efit of U.S. provisional application Nca.
60l81'.6,t)6-3r fil.ed 1tiiic, 2:33), 2006. Additionally, this application is a c:a,~ntintÃation-in-part of U.S. Serial No. 11,491,829, filed Tti1y 24, 2006, which claims priority to US.
provisional application No. 60/ 70(,903)õ filed JLi1y 12, 2005. `l".Iie etitirc co-ntcnts of these applica-ions ai-c incorporatezl,by i-cfei`eia.ce, hereirl.
C,Oi"Ett:NMF;N`!' RIGHTS
10tl02] This researc.la was stapported in part by tE-ie Nat:iotial Institutes of:1~-le'-flth (NIH P50 CA 10-3 ) 1. 75}. "I'lae gz ~'vernment of ttae Unitcd States mav have rights to this i~~~entic?ll.
FIELD OF THE INVENTION
10003] The prescrat invention .rc.lat.c.s Ãc) a iaiic.ro#~Ãbric~~ted nara.o-or Micro-sca1e particle for encapsulation and deliv~.~ry of i-naterials or substances itic:.tticl.ing, l:aLIt 1-10t limited to, bic31ogica1. incdia incluti.ir~g cells, pharinac.e::cabc:al agcnts, cornpositicarIs, dra.igz, ti;;sue, ge[s and po[y.mcrs contained vvitbiai the laci_rticle, wit1l subsequent release of the tkacrapeutic materials in situ, ri-iethods of aYaakitag the particle atad arletliocis of uszrag the part.icl.e in in. vivo or i~~ 'vitrca applications.
1;3At"11,,.GROUl~lD OF Tt-JE INVENTION
10004] Tn recent ~~~ms, advances in regenerative meclic:.-ine bave inspired therapies targeted at the ccllLilar lcvcl. TI-icse tlaerapies seek to implant cells or ceilLilar clListcrs>
manipulate cellular pat~~~~ays: aia.d target the delivery of drugs. For exwiaple, a wide range of cell liiae;s laztvc bec:ra csicloyed wit.liii1 semipermeable iiÃid biocornpatible irÃ-u-nobil:izaÃion devices that control the bidirectional diffijsiorà oi rÃ-iolectÃIes and cell release (R. P. LaÃira, J. L. Hayes, W. L. Chick, Nat.. :BioÃechnol. 14, 1107 (1996); t3.
Drive, P. M E4errÃatÃder, A. R Gascon, M. ~~ariÃ.aa, J. L. 1?edr-az, Trends irÃ
13iot.ec;.}Ãno1. 20, 382 (2i30'.); N. E. SÃnÃpson, S. C. Ciratit, S. J.
131~ckbalÃd. I.
Constantinidis, Bioniate:ria1s 24, 4941 (2-003.).). Concurrent advances in microtechnology baare revolutionized medicine; as Ã~ew implaÃ~table devices, ziÃicroarrays, biocapsÃ:ales aÃid mia;rcÃprcÃbes -a~~~ developed. "('tiese dev.ices have tacilit:a.ted ce[lular encapsulation, on-demand drLÃg release, and early diagnosis of dis~~~es, (:1_ T. Santini, M:_ J. Cima, R. Langer. Nature 397, 33-i (1999)s J.
Kost, R.
Lan4,~eÃ-, Adv, Drug DeliveÃy ~~ev. 6, 19 (~ 99t ); L .Leoni, '1'. A. Desai, Adv, Drug Delivery Rev. 56, 21 1 (2004}; B Ziaie, A. :[3aidi. M. Lei; Y. tFiI, R_ A.
Sieget, Adv.
Drug Delivery .(~ev. 56, I45 (2{304); A_ Desai, r! _ Wes"t,.M. Cotien, 't'.
:L3oiaÃsski, A.
Rampersaud, A.dar. Drug Deliveay :E' wv. -56, 166:( (2004); J. T. SaiitiÃii, A. C. R.ichards, R. Se}Ãeidt, .-N4. J. C'i~~~.-za, R. 1n.aÃigea~, Angew. C'beaYÃ. 39, 2396 (21000); L. .FireÃ~ian. E
Mahaji7as E. Broide, M. Shapiro, -[__. Fieli, A. 4terrlber; Y. Ke~lielmati. E.
Scapa, Gut 52. 390 ('U0)3~3_ In ~:Ã~ntr~.st to ~~cAi~raneric, l~~rdr~~gii, and ,cil-ge( based prw~cesses that have been tisÃ;ci for encapsulation and delivoryx coi Ãvet Ãtiona1 silicon (Si) based microfabrication has higb reproducibility, provides ÃiiechaÃiical and cliemical stability, a.iid allows the iiiccarpora.tiota of electrcÃiiie and optical modules witllizl the device, therf'.b~' facilitating wireless telemetry, remote activation and C:omTllÃ,iTl#cat#oll, in vivo.
Howevc:r. Si based microfal~iricdttion is inIÃerently a twti dimen<i~.~tia( (21)j process and it is extremely diffictilt to fabricate three-dimensional OD} systeÃ-ris using conventional microfabrication (M. Madou, Fundamentals of Nficrofabrication (CR.C, Boca Ratc}n, FL, 1997)), ~ 3D medical elevicÃ; has several advantages over its 2C3 c::ounier~.~saxt; (a) :-Ã
laqger~ emerna1 stirl`acc area to volumo ratio, thereby maxiiiiizilig ititeractions with tlie surrounding medium, and providiÃig space to moLrtit di.tTerent diagnostic or deliveÃ-y modules, (b) a finite volume a1k)~,ving encapsulation of cells and dnigs, and (c) a gc;ometÃ)~ that reduces t(ie: chances of the device being Ã;triclesiral.~1v lodged in the bt~~iy-1(1{}051 .I.n orae aspect of the larescrit iriveritjon, biocontainers (i.e., boxes, I-iollow particles) have becti fabricated by a strategy that conYbizies the advantages ot tfirce-dirr~ensiorraliiv with the desirable a5peÃ:.ts of Si-based microfabrication to facilitate the delivery of i:tier~apeLatic k(gents isi sitii. For ~xampie, the eoritairier-s are loaded rith ztricrÃ~~eads or cells e:rnbetlded in a gel, arid thus Ã:mi be used either in coqji.rnction with preserit dav immobilizatiori svstents Ã.ised, in cell eracapsÃ.l1atioii technolog, ~~, or tl~e~;~ c~z~
be used zndqendently. In a:riot.lier aspect, the biocontainers also caii be used fcar encapsulation o1':fiunctinnal cells witlairi the por-oras containers ior in vitro arid M vivo release of therapeutic agents with or wit(ioLit amma,rnosuppression. For exarnple, the cor-ltainers caÃa be used for ei-aeapsulatiori atid deiiveryot insulirx se-cretirxg cells for-iniplant-ation in patients wviÃh diabetes, for placing tLsmor innocula in ari:iriial riiodels where, e.onstra.iruna, cells ~,viÃltin a small re~fio.it is necessary, and ior-de.liver~, of tuaictioraal iietÃronal ('C; l2 cells. :I rt some embodiments, tlie faces of the container are patterned witli riii<:,roscale per-l:or-atiozts, allÃ3wint; control over per-#t-Lsiort and r-el~zise ol its contents with the sug-rortndirio; rYiecliLim. `I'be advarita~~;cÃ:uy attfibutes of the contaiiicrs are a parallel fabricat:ior~ process with vrwrsatilit~' in sizes and shapes; precise and monoel.i~pea=se strÃfirc:e porosity; and the ability for renwne guidance iisÃr~g magnetic ield.s. In another aspect, the coiitairzer-s of ttie present invention are easily detected a.rid non-invasively tracked i-ising CortvenÃional rt~agrletic resonance irnagin g (1LI~'~Iiand.
do not require the presence of a c;o-ntr-ast ~.~~ent, S'Li M_MARY OF THE INVENTION
1000G] The present iÃiveÃit:icÃ~i provides riaÃxoscaIe or microscale particles for encapsulation and delivery of materials or substances, xncludiÃxg, but fic?t.
l:izniÃed. to, cells, clrLigs, tissue, g;elc aÃ-id polymers cotitairied within the particle, with stibse~.~LÃeÃ-lt re:lea.qe of the therapeutic materials in situ, methods of #ab.ricatÃÃx~~; the particle by foldfiu-) a''D oreceÃ.rsor iÃito tkÃe ;3D parÃicle> arid tiae tÃse of the particle in in-vivo or in-vitro applica.tioris. In oÃ~~ ~~iibodi~iieÃit of the preserit iriventicÃtÃ; a three-di.metÃsiort-Ãat particle cÃ:Ãmlarises a mLiltitLide of Ã~~vowditnensioÃÃa1 faces itlat form a hollow, polyhedral shape a.iid containing a IillabIe center chanibe.r, whereÃÃl a.
size of tlle particle is niicrosc.ale or tiwiost.alc;. In wÃotlÃcr emliodimcnt the two-+:iim.ensiona1 faces of the particle are l.^~attemed wit.li 1.~eÃfibrations or pcares. .1n another embodiment, the perforations or pores are created photolitliograpliically . 1Ãx another embodiment, the perttarat.ions or pc?res have a. size from abotit tl. 'I nm to aboi~t 100 mic::rcÃns. 1:n another embcadim.entr the particle is fabricated troni at least one material selected I'rom. the group consistiÃig of a metal: a. polymer, a glass, a semiconductor, a11 insulator, and combinations t:}ie~~eof In another embodiii-aeÃat, the metal is copper or nickel. tn a.Ãiot.}ier embodiment, the particle is a Faraday t.a~,Te. In aiiother onihod:inient the particle is coated wit[i abiocompatible materia[. In arivlhgr embodiment, IEle biocompatible material is a metal, a polyÃxier, or a combiriat:icari thereot.
iÃ~ another iriil7odiriic:iit, the filla:ble: center chamber of the pa.rliclo is filled with at least one sÃ.ebstance comprising coiitents of the pa.rticle. In another einb~.~diinÃ;nt, perforations or poÃ-," in the iwo-dimeniioÃia1 faces of'the Pa.Ã-tsc.le allow re'lea.se of the contents of the particle. In another embc3dimentM at least one substance is ~a therapeutic ageÃit. In another embodiment, the the.rapeutic ageait is selected froÃn the {;~rULÃP
C0D;;iShÃ16Jr OI' a cell, a plÃannac.etaic.al. agent, a, c:ora-:tpos-ticÃn, a tiwstÃe, a. gel, arÃd a polyrnt,r. In another embodiment, t}ie particle is admlfl-iistered to a sutject and location of'ihe parfic.le HI the S1.1bjecà is non-invasively tracked by i31agi1ek rOSC1T1ai1t:-C i111a<q?ing.
I17. iÃ.I7.C3t.l7.Gr erÃabc.>dirnent, ttae particle is iniaged with negative coÃ-it.ra4t retiÃtive to back,F{rcawid or positive cLiÃ:ftl"cÃ.St relative to background.
100071 '~(`hÃ: present invention also provides a niethod of .fa,f3.rie~~Iti~~~
~1 il~~Oe-dinieÃ7sional particle comlarising a Ãiiiilt:it~ide of t~.vowdiaiensional faces that l`orÃ~~ a hollow polyhedral shape aiid containing a. fillable cetite-r= chamber, the method cornprÃsing tlie steps: (~~) fabricating a rnultatuele of Ãe.vo dimensionat faces; #.b1 patterning the fabricated ÃwoTMd'amerasioÃia1 faces; (e) patterning at least one hin4}e (ari the pattemed two ciimetisional face to tori-n a Iiiriged ed(;e; (d) J'oinitig a hiÃigecl ed~.~e of a first laatterried two dÃ'iiicrisiozial face to a hinged edgge of a second patterned t"i-o dimensional #ace, to torin a IiiÃia;ed JoiÃat; (e) repeating step (d) to form a two dimensiona.l precursor template having hinged joiiits between ad~acent two ~imensioÃ-tal taces; (t") lÃiluefiying, the ti'txiges of the two-dimensional temp1a.te using heat; <i.Ã7Ã1 {~} selt:r-asceiiif}ling the threercd:iÃvierisaonaI ~~t-ticie_ In another eÃrtbodi~~~~i-it, tfic hinges ol=_ ~tep (e) of the method comprise a material fhat can be liquefied. .(.ti a.iiot:lier ciiilaodimejit, the material is a. sold.er;. a. metallic allov, a polymer or a glass. In anot:h.er emlaodzmeiit, step (a) of the Ãiiei:liod 1~irÃber coÃTrprises the steps (:i) spiÃiriiiig a sacrificial fili-ii ori a substrate to 1 riYi a tirst Iaver, (ii) ~averÃng a conductive secoriÃl layer on the first layer; aaid (iii) patterning the layered st7bstrat~ by photolithography.
l:ii another embodiment, the particle has a size that is microscale or nanoscale. I~
another embodimeiiÃ, in ste~., (b) of the iiiethodt the ti.vti-dimensicrna1 faces are patterned witb pertoratioiis or pores. The perforations or pores are created photolithtagr;:tphica11~,. In another e;t7~~od:imentõ the perforations or pores have a size frtani about C).'1 iiin tca about 100 microns. In anotber einb~.~di~nent, the particle is a Faraday 100081 The presetit invention tt.irther provides a method of imagita.gy a tliree-dimcansi.ontil pat-tic:lc comprising a MtlltitLac9e of two-dimenwi~~ial fiaces that form a la~.~ilow polylicdra1 s[ial.~e aiicl coiitairiigzg a filla.blo center c~amber that Eias beon implanted into ~subjject, t~~e method, comprisiaxg, the steps nf` (i) loading the fi1:la61e center chamber of the paa~ticle with at least one substance to form a loaded particle; (ii) admiaiste:ring the loaded particle to the su~ae.ct; aiid (iii) noninvasively tracking the particle of step (ii) in the srIbAjec:t by magnetsc resonance imaging. In an.offier erribc>dinicia.Ã, perforations or pores :ir.a the bi-v'o-c.linieiiSional Eaces of the particle allow release (yl"tkae subcÃ.a.tac.e in the fillable c.criter ch.a.iTibcr. i~~
another etaiboditaaetai:. the at least on.e sub,,;iance of step (i) is ~thea~apeutic agerit. In atiotl~~~~
emliodymet5t,, the therapeutic a.gcnl is selected from the 4~rOa:ap corlsisting of a ce11, a lsharÃnaceLstical agent, a composition, a tMue, a <=,el, and a polya~er.
BRIEF DESCR[PTION OF THE FIGU RES
100091 Figure 1 is asclaeraaatic diagraiYi of the process flow used tot.a.bricate the :')D containers o1'tlae precetit invenÃz011.
100101 Figure 2: (A) Optical inaage showing a collection of containers. (B-DI) Optical and Scanning c(c;c:trosi microscopy (SE.NNI) images o:l'micrc~pattemed cuntailiers at ditTererct st~~~s of the fabrication proeess; (B) tlie 2L3 prectirsor witli electrodeposited 1`aces: (0 the precursor wiala faces and lainges, and (D) the folded container.
1Ã1011 1 -Figure 3: (A) ~ENNI iniage of a laolltiw; opGiiaf-tced coiitairier.
(B) SEM
image cif a cojataiia.er loaded u;itla glass ixaicroboads. (C) Optical image of a biocoÃataiiier loaded with MDAMB-~? j I. breast cancer cells embeclclecl than extra-c.cl:lular matrix ÃEC.N-11 gel. fl_7;y Fielea.se of the cells by immersioti of the container in war~i-i cell culture medium. 4E;1 Optical image of a container loaded with a ceI1-E~."M.-ag,-arose srispetision stained with the tltflorescent cell viability staiÃa>
CalceÃn-A:M. (F) Release of the viable cells from the container on itarnc.rsdon in warm cell culttire incr~ium.
10012.1 Figure 4: i4I_RI im~~~~~s of an ~peti faced (A;) noii-magnotÃc: CLi c.oiitainer a~id (B) ferromagnetic Ni coÃataiiicr. (C-D) 1~'itiitc e1~~~-nent simia(atÃon rewLilt; of the Mar rnagnet.ic field in the region of a L"Lgcontainer, iii the (C) xy aTid (D) yz ceia.t.ral planes. Ttic excitation cÃariaprised a linear polar.izecl 500 Ml-lz plane wave ofl V.'ryi, with tlic E. atid H fields in the z and y direction re;;pecrively. Tlie magnetic field cl.istoTtioias aaad the shielding effect caLaserl by t1-le wire frame are eviclent..
100131 Figure 5: M:I;, trta.ck.irag of a coniainer in a flLiiclic chaiiiie;t.
MR inlag ge;;; cal the container at tliftert.nt tinae points taken tÃ:iicler pressure drivc:n flow of'the fl.uitl.
1 (1{}l41 Figure 6: Optical a~id Sl=:~M i.7nzr_ges of the tl-iree steps rrsed to fabricate mic.ropattemed boxes. "i'he boxes sliowÃi iiavc approxiaratc dirTletrsioris of rxricr- ns. Frc~i-ti left to Ã=iglit- (a) The faces wer=e patterned using..-photolithography atid e:Iectrodepoaition; (b) Solder- hinges were ~.Ii~Y.t~ed rel~.tive to the faces usirx4;
phoÃolitho,graphy, etching and eIectrodepo~ifion; and (c) the 2D precursor was-lifted off Ã1~e wafer upori dissolutiari of a sacrificial (aver. When t.}ie 2D
precr.rà Sor was heated above the riie;,titig poirit of the solder, the siructr:rrc folded ilito a 3:13 cubic box [B.
CJimi et a1. voi. 7, p. 341r343, 200-5_I.
1001.51 Figure 7: Images of sorne defect modes observed: tt the solder }lei(Yht is not optiaiiiz~ed (A) uÃ-aderfolded or (B) overfolded boxes are obser-ved. (C) Incomplete etc.:hints of the seed laver usriallv results in faces that cannot fold (i841"),, because they are fu~ed together witii the seed Iaycr-100161 Figure 8: SEN1 arid optical images of boxes tilled wit~ (A & B) I?iuroziÃc hydrogel aiid (:') MUA-'-9:13-23 l breast cazieer cells embedded in e:xtracellular matrix (ECM) ge1. (D) =1'he cells could be rcleased from the box b;: pulsatile z~~~:itation in cell tt-redia.
100171 FirÃit=e 9: (A&B) Optical ii ~agos of 2D c(iÃl; fabricated tisir7g photolithography. By passing ct.rrrent thr-or,rgh tiie coils it is possible to generate a magrietic field. (C) The microbr~~ is placed alorig t..fae central axis of the coil in order to i.nduc:iivclv heat the bc3x, 100(8] FigÃire 10: Release of dye from a lo-idc;d. box r.apon heating.
1.00i91 Figtic=e 11: a) A scaaining electrori microscope irnage of an empty container. The containers were three-dimensional (31)) porous cLrbeS with a length of approximately 200 mm and a voILrmc of 8 nL. b) Ari optical microscope image of a coritai~ier loaded with a dye-soa-k-cd pl~roriic ge1, cf A scliematic diagram of t.lie ~Nperimental set-Ã.rp used to facilitate wireless micr~~ascaIe chemical engineering (Ãlot drawri to scale). Ccgr7tainers were manipuIfrted uSing a magnetic stylus (not Shwvn) and.
the contents of specific c~titaizie;rs were released by directiiig an RF
source tciwardS the container. In tbc; schc;matic representation, chemical Y is releia ;d from a specific conttzine.r; c;liciia:ical Ytl.aeri reac:ts wi~~~i chenaical N. in t.lic 4urro-und.ing r~-iediui-ii f~ca t`orni pt-nd uct I.
100201 Figtire 12: Optical imkges showing the reriiote controlled, spatially ]ocalized microkabgication witliin a cap:illary_ 'I'wo i-iiaci-owi.r~s (l and 21) were embedded within a macrnl='abric,atecl c.apallw-y (ca. l mm in diameter and 1.5 cm in length) aiid the capillar,v was aligned on. top of a ZD microc.oil, a, b) First, a container filled wit:li piriroiiic and soaked with the elicmÃcai sensitizer was guided iiito the cap:i11a.ry to t}ie site of the gap within wai-c 1usant~~~ a magnetic stylus, c) "l'~ie, cllemical wnsftizer was released bv re~inotefv heatiax, the sensitizer-soaked pluroÃiic gel that was encapsulated WithÃn the coiit.~~iier. '1'-(iis heating was achieved witb the 2D R:l?=' coil.
After sensitizing the gap, the first c.otii:aiÃier was removed, ~~econd colit.aii-icÃ- lvas guided to the same gap in microwire I. -i.tad t:fae actMator WIS released by heating the pluronic gel renioÃely, d) Atter activation, ttie second container was also removed. e}
The capillary was tlicii filuslicd with a commercial electrn1eSs copper-plating Solutiori;
chernica1 reduction (bubbles o1'thc: hydro4,en gas, a byproduct iti tt~e reaction, cari be seen) of copper sLilt atc to metallic copper, occtirred at the gap within microwire I. t) Coppe.r was deposi.ted otily in tl-iz gap $.~etwe~~i mic.rowire I, ~io c(il?per was depositc d.
in the ga~., f niiiierowire 2.
10021] Figure 13: Cell-viability asSe~smeiat by five/deacl fluorescent imaging of ca1c;eiai AM and ethidium homodimer-1, both released rei-note1~~ from the containers. a, b) Confocal iiua.gcs of the local release of the live/dead st-~n tci L929 inouse tibroblast cells. No red cells were observed, thus indicating tio nec.rkotic cell death during the aelease_ a) Transmitted light differential interference contrast (DIC) images sho-wing both the cells and the coiit-ainer. l;i) F(~~oresc:enr image showing on13=
localized cell sÃa.ining, 100*2 :1 Figtic=e 14: (A) Optical image of the color c.hang;e observed on a temperature indicator label placed under a nanoliter container, expos'ed to RF
raciiation. The color c l}agige occurs only under the container ~~owing that the heating is local. (.B).A plot oI'the temperature rneastared using t.lic coior-indicator Iabcl vs. the i ticicicnt RF power.
100:231 Figure 15: f_'csnfoca1 tiric:roseopy ir7rage of a tiatioliier c:orrtairicr loaded witNÃl?Nl:l?;1.i-zr gel soaked w.ith 1.1LEIDEAD<"~ assaw. 'i`1-le crpcrin-lent iotr?wed the test proceclÃÃres, v~=ith tlrc exception that the RF was not turned oti. --t'1Ãc absence of cell stainin4,~ surrounding the container (corÃxpare witb Figure 13) demcynstr-atcs no dzsccrr7a.lal.e c}Ãcrriica.1 release in the abscrrce of the RF radiation trigger.
fUtl24J Figure 16=. C 'orrrparison of finite simÃilaiio rand cxpcrimerri.al results for the self-assembly process, (A)'I~op view (drawrr to scale) wittr dimensions of the faces ~~id gap widt.}rs of ti-ae'?`D template used t.oselt-assemble t.lre cube. (B) Side v:ie-%.~~ of two ac.jaccrrt faces nf the crucÃt`orÃir (as tabricai:cd),with variables Ãased in tl-ic liriitc dement sirxiulatiorr. (C) Side view ol'at~iacent tacesat the oriyct o#-rci~~Nv r>f'tlie toldin }ainge. (L)-0 Firiite element snapshots sbowirrg (~~) urrcle~~folded.
(E) right-atigIe 1o1ded, aiid (F) overfolded fiaces. (t -I) Optical nricroscope iÃ-zrages of cxpcrirne.artall;; fabricated 200P.m ciibes exlribifing the Lsrlclcrfoided, right-angle folded, and overt olded faces. Note: Fig t.B-F are not drawn to scale in order to illustrate important (iiz-ncti5iosis.
100251 Fi~~tre 17: SimÃÃ1aÃiogi results of ti~e depciiclc~ice of the fold angle on solder vc-i1uriie The restilts deÃ~~~~iistrate that folding angle ca:ir be precisely eri;;irrecred by controlling the s(?Ider- vvIÃimc at the hinge.
100261 FigÃire 18; Normalized total cncqgy curves (finite c(cniciit simulations) plotted as a fiirictiorr of fold ang;fe for faces with Ieragt.hs r-arrgin:;
frorn 6mrn to 50 n ni.
The curves show that folding is spontaneous at sma.ll size scales with stable rafflirna.
As the scale increases, gravitational forces increase and folding is no longer sporrtanc~.~us (iiiitial slope cliariges t'rorrz riegative to positive) and there is rro minima preserit a:t 6 mm.
10027] Figure 19: (A) An optical image showing tiee standing polyhedra fabricated (c.xpc.rimeiita( rc.sÃilts) witl~ a. wide range of sizes all the ~~~av from '~~im to (B) 15 tÃni, and with ditTt.re;nt shapes e.g. (C) A ;;tlr.tare pyraniid.
100281 Figure 20: (A) Optical ii-iizt~,se of cubes With a .ratigÃ; Of StZCti I`zsrTtic-ld in lar~.=ertunilaers. {'1::3} lonri-ied in iniage of the otitlitied re~~iorl in (rl) feai.Ãtt=in~~ I00 pm cubes siitÃtic, oti top of and anion~..9 500 1mi ecgbes.
f00291Figure 21: Sync3pa.iS ol'toiding process.
100301 Figure 22: l~) Optical atid SE'~~~ images s17o-,.-ving the dÃtl'eretit steps (tlie photolitho4,~ral~l~.iÃ:..aJ1~' fabricated '?l) template, Ãe~;ist:r-~; of solder ItiÃ~tges and the folded -:D s[:rttcttire} in the fabrication of acttbic. container with one t7peÃi face. SEM images ot a(b) ct:tbic cotitainer ~vitb all opeti flaces, (e) pyrannidal fr-uStut-ti, (d) sqttarc pyramid with ati open face oii the bottom, (e-g) Optical image of multiple containerS
of dltTereait shapes demo:istra:tinia tl-ae paraltel tabrication strategy> {h=h.}
SEM itx~ages of cÃtbic cotitaitiers witl-i monodisperse pore sizes of (h, j) 5 atic.t=ons atid (;r; k) 3 m:icrÃ?tls.
10031] Figure 23: Optical images of chcralcal release from containers ( a) Spatially isotropic release of a dye from a container witb iclentiÃ:,a.l I?Ãarositv, oÃi all faces (la) AniSotropic release of a dve I'rozii a cÃ3ntaztaer- with aÃ-tisot-ropie porosiiy (five faces with ati array of 5 iiiic.roÃ-i porcs; ilic stxih face has a 160 i-tiicr-oti sized pore). (c) An e\arnplc of a remotely guided spatially controlled chetnical rÃ:;tction. The lettÃ:r G (for the Gracias Lab) was for-nie.t.i by the direct writirig of pheaiolphtlialein in an alkaline water---glycerÃ.~[ meclit.tm.
10032] Figure 24: Spatially cotitrolled cliemicai reac.tions. between multiple containers. (a-c) Reaction of copper sulfate and potas;ait,tm hydroxide in. an aqueous inÃ;Ãlit.tm resulting in the tortZ~ation of copper hydroxide along the central llrie between the Ã:..ontainers. (d-f) The reactlon of phetiolplatlialeiti (diffusing caÃtt of the two bottom containers) and potassium hydroxide (diffusing ottt of t.lie top cÃ}ta.ta:iner) in an aqt.tec=t.ts medi utii.
DETAILED DESCRIPTION OF TRE INVENTI+EON' 100331 The terms "hollow pat-trt:Ie." "box," ::contasner" and s.b1C}coI7tcltl7et`õ zEre used it1tf'.rcl1ai7~~eably f3ereii7 to i3lLili3 a three-dimensional C1bjjGct, i.c.., a rec:c;ptacle, with a hc.3i1mv inter'i.or- or an ititt:rlor capable of containirl_g sttbate~i-ic;e;;.
100341 I`lÃe term <;colloid'R or .:colioida(" as rase(i herein re::f-er-s to a substance tiÃrÃde Lap of asy5Ãem of partic[es dispersed in a c01Ãtia-ILiOr.aS r-freeliÃ:rm.
100351 Materials carà react quite difTeÃ-eÃitly iÃi the presence oI'aÃ-i extemal nia:grÃe't.Ãe tield. 'I'kiei.r reactic~ri is depe.rident orà a rart.inber of factors, including, but rioi: liÃYiited to, the ntaterial',, molecular structure, its, ator-rtie strLactÃ.rÃ-e, aÃid the Ãtet mao;Ãietic field associated witli the at.oms, Most materialS can be classified as fot`romagnetie, diamagnetic, or paraiii agri eti C.
1U03~"iJ The lerÃ-ii "diaÃnagnetic"' as used laer-ein refers to rYiate.rials having a very weak l'cirtti of magnetism exbibited only in t.lae presence of aÃx external magnetic field, which is tl-le result of chaÃtges irà the orbital motion ol electrons due to the exterrÃal rnagnetic t:ield. itidticed nia";ÃleÃ:ic r'laomerat. ÃÃi a diamagnetic mai:er-ial is very small and in a directioti opposite to that of t:lae applied lield_ 1-Nartipies of diama~.t1dic materials include, but are Ãiot limited, Ão, ct~laper, silver and gold.
1.0037:1 The terrn "1earornkgrteÃzc.,, refers to tnaÃeria.ls t iavirlg large and pca5itive Susceptibility to aii external riia(sÃietie field. Fer-romag-rietic rim:teÃ-izt.ls liave soÃxÃe ÃÃnpairec.l electrons so their atoms have a net ri-aagÃaitie moment. They e<bibit a Stront), attraction to t~~agnÃ;tic iÃ;lds and are able to retain their magnetic properties after the external field Iias beeri removed. Examples of firrr~.~magrietic rnateria.ls includel but are ntit 1'imited to, iroti: riickel and cz ~balt.
10038.1 The ter-rai "param~~~r-ictic: refers to materials lravi~ig &Ã small and positive sÃ.ÃSc:c;ptil.~ility to ni.a.gnetic fields, wliicIÃ are slightly att.racted by a rnagneiic fiel&
]'a:ratnawictic materials do not retaiii magnetic pr-opeÃties when the ext~.rÃtal tzeld ~is r~~noved. These paramagnetic properties are, clÃ.7e to the presence of sor~e unpaired electrons and the realignment of the electron orbits Carr.secl by t1ie extemÃ.-r~ magnetic field. ExaÃzit.~lcs of'paramagnet:ic rriaterialS inclrik, t~~Ã.Ãt are Ãi~.~t limited to, magnesium, molybdenum, and litlaia.rm, 10039] The terÃai "Faraday cag&' as t.r~ed hereirr refc.;.rS to a:ri enclosure designed to block the e#Tects of an ele.eiric field, wliile a11~),~ving free passage to magnetic fields.
(Sef E. M. PtÃreel1, Electricity and Magnetism, Berkele-y Physics CoÃ.ir ;
VolLÃ~ie 22 (N-l:cCfr-aw 1:=[ill, MA, 1985)B. SÃic}i an enclosure also is called a Fa.raday shield, Faraday Shie:[ding, Faraday ccrecsi; Faraday electrostatic sliie1dõ or shielded room.
100401 The tet=iii "gel" as used licreii-i refers to an apparently solid, jellvtike materia.l fnrnied :fi:rÃ~~~i a colloidal solution. By weight, 4e(s are niostly licli.-Eid. yet they behave like Snlirls, I"_[ie tenii "solution" refers to a homogeneous riiixture of one or more substances (tlie soleites) diss1 ~(ved in another substance (the Sc~lverit).
100411 '~('lic term "izidriÃ;tivc heating" as used bereiti refers to Ãtle process of heating a znetal object by electrognkgnetic iÃiduc.t-iori, where eddy currents ar-e gei~erated within the metal and resistance leads tc) Joule }ioatiiaz ~~t tbe metal. An induction heater (far ~~~ v proc.ess) coÃ-asists of an electrcima<wnet., throÃi<wh whiÃ:.1~ a hiwh-frecluency Alternating Current (AC) is passed. Heat mav also be (5enerated bw magnetic I-iyst:eresis losses.
10042] The teÃ'Ãn "magnetic field" as: L&scd Iicr~~ii r~~ers, tc) the re~ioti lri 5pac=~
surrounding a magnetic bodv or erÃtit:y; stÃc.l~ as a pertrianent magnet or a ccyiiduc-tar carrying a ctirreÃit:, w(iere ati appreciable magnetic force is preSCtiÃ.
Suc~~ a field is rcpresezit:cd bvinagriclic lisies of force. In an ciectromagnctÃc field, for exaiYipte, the magnetic field is perpendicular to the e(ectaacal field.
100431 The tÃ;rni "z~~~iielic field strength" or "magnetic field inten,ityõ
(<:ff') refers ÃÃ.~ tlie iiiteiisity of a ina.gnetic field at a given pÃ.~iiit. Mag-netÃc field streti:;t}i is, a vector qtÃatitity usually expressed in amperes per meter or in oersteds.
10044.1 The term "magnetic reso-nat7c:c imaging'. or "MRI.", refers to a noninvasive imaging technique that ~~ses the interaction bet~vee.n radio frequency pulses, a strong magnetic field, and an subject to construct images iti slices/planes from the nuclear magnetie resonance (NTMR) signal obtained fic}m the hydrogen atc}inS inside the suNect. The priiicip(Ã; behind all MRI is the resonance eqttatson#
100451 t1=:; B+, ( ~quation 1) 100461 which shows that the resonance frequency u of a spin is proportional to the n~~~~~~ietic field B~; it is experiencing, w~iere y is the gyromagnciic ratio.
100471 As used bc.rei.rr, the tÃ;rni <;mic:ro;Ã,a:lc" refers to par`tiÃ,lcs -tliat rrreaswret`rcarn about Ijinr or I x i C)'"; meters to about 999 l.irn in at least orrc dimerrsiorr. As used beÃ-c:iti the ter-iii .:.nanoscale" refers to particles tkiat measure trort-i i.l,oÃrt 1 rianorrietcr- 0r=
1x 1 F)`9 zrrcte.a s to about 999 rraÃroinctcrs.
100481 The term "ma gÃretic field ~radient" refers to a variation in the ma4}rletic field with respect to position, A one-dimensional mag-neiic field gradient is a variation with r-espect tci oric direÃ,tioÃi, while a two-dimcnsiozral ggradiezrt is a variation witli respect to two dircctions. `I'1-ic rr-iosi Lisct=ul type of g;radieÃrt in rnapetic resonance imatyirrg; is s a c~~ii~- dimensional liaxea:i- magnctic t~eld gÃ-ad.ient. A one-dimensional magnetic field gradierrt along the x axis 'Lrr a rxragrretic field, B,, indicates that the zrragrretic field is increasing in the x direction. I'lie symbols for a rnag;neÃic field gradient in t}ie x, v, aiid z directiorts are G:;, G and G.
100491 In physics, the term "riiagnctic rr~omerit" or .<dipole moment" refers to the pole str-ength. cit a arragrictic sour-ce mttltiplied by the clistarice betweerr the poles (p pd), arrd is a measure of the sÃ.reÃatYth of the niagnctic source. The rriao:rietic moment in a magnetic field is a mea.stire of the magnetic t1ux set LÃp by gyration of an clectroii chrirge in a mr~ggnz.tgc ticld.
100501 The terrii "micrÃ.~paÃtern" or .`r7ticropatterrreel" 'ati tÃsed lrerein refers to ar1y arbitr ar-v, two-dimensional pattern haviri<w microscale feat.tiÃ-es. The term nawpatternõ
or "nanopatterned" as uscd herein refers tc4 ~nv. arbitrary nsro-dimeÃisÃonal pattern l`Ãaving microscale teattzrc;s. Akcordirig to the present invcnti<Tn, the particles are paÃtemec1 with 1~erforatioÃrs c~~- pores r-aargirrg in size from abntit 0.1 niri to about 1.00 ml ct'ons.
1Ã1051 I The term "oscillatig~g magmetic field" or "oscillatory maynetsc field" r~~fers to a magrzetic field that pe-i-iod-ica11y increases arid decreases its irrtenwity, rlr, or wlricli othenvise varies over time.
10052] The particles of t~ic present invention may be in any polybÃ:drai shape. The term "polyhedral" as tisc;d lie.reiri refers to of or relating to or resembling a l,olyhc._dron.
The ierin "pulyhedron" refers tc) a three dimensional ol~ject boLindcÃ~ by plane po1ygoni or faces. T}.ae terna "polygon" refers to a mtiitisiticti <;cornet.ric figure that is bnui7ci bv matiy straight iiÃ-ics; such as a triangle, a sriuare, a pcrita~~~ri, a ~ie:~a~~~ri, a i~epta4ron., an octagon, atic{ the liike. FoÃ- cxatnpic, the particles oi't~ie-pr~s-etit i~~~~~~itiori mav be a cLibe or a tetra.liedrai.
10053] The te:rin "radio freqLa~iic.y" as used beaeiii refers to a fietitieiicy or interval of trequeiicies witiaiii the elect:roaxa~gnetic spect:runa Lised fibr com.munÃcatÃons, usually dcfiiicci as :~paiiriiiig fi-oiii aboui 3 k.Hz to about 300 t.i=HZ, w1licfi c~~tTesptlrtlds to wavclenubs of atjotit: 100 km to about I mÃii rcspectively, [0054] The term "radio trequeiicy tag" as tised herein includes radio frequency icÃeait'tfj cation (IR~~ID) tag:=. Radio-frequency identification (RFID) is aii azitomatic identification method, relying oii storing and rcmotely retrievi~ig data Easin;,~ devices called RF:(:D t~w=s, Ati :ft_[tID tag c.ati be attached to or incorporated into an object i=or the pur-pose of id~iiÃificatioii Lisiiig i-adit-~ waves. RFID ta~.s come in three geÃ~,erai 1,)wsrre, serrri -pwssive (also known as or crctive, Passive tags rec{u:irc iio iriter~ia( power soiirc.e, whereas s~i-iiiwpassive and active tags require a pt~~vea source, usually a small battery.
[0055] The tenii. "resistance" refers to a measure of the ~ie(zrec to wi`iicb an object opposes the passage of aii eicctric cLirrent as re~.~resclited by tile equation, RmmVs/I, where R is the resistance of the object (usuaIly measured in ca}iins, equivalent to J
s1~"}; V is the potential differe;nc~ across t.iic c3bjjec:tx usually measured in volts, and I is the curreiit passing thr.c~~igii the objjcc:t, usually measurc;d. in amperes).
1.005G1 The presence of a~iv sLibsta.iice in a inagnetic field alters that field to sc~~iie cxtent. The temi "susceptibility ettecx" rcfirrs to the degree to which a sLibstance't inherent magnetic moment prodtacc s polarization wheii placed in a ma:,rtctic tieid.
100571 ft'Iic tcrins "two-dimensional" or ::2I)'.' are Lised interchangeably herein to refer to a figure, ol~j ect or area that }ias i1eight and width, but no depth, atid is therefore t7at or planar.
10058_1 The terms "tbrcc-dimc.nsionaI;~ or "3D" are used interchangeably herein to refer to a tigtire, cgb_jtact or area that bas height, width, and ~lepth.
100591 I`lrÃ: particles of the prc5eait i.7averi-tiÃxn are fabricated using aÃ. least caric material selected :firÃ~ni tkie M,rotip consisting of a rTlctal 0-neaning aÃ-r clenicnt that is solid, lias a rxtet.aIlic lÃa-,terõ is malleable and dtrc.til~, aticl conducts both liei:t arid e1cctricity), a poI;;iiier, a glass (meaning a brit-tle transparent solid witfl iÃreplar atomic structure), a Seniicoridr.rctor (meaning ari e:leriieritõ ~swc1i as silicon, that is inÃerrnediate in eleÃ:.trical conductivity bet.weeri cor-rdLrctorS and insr.rlat(yrs, through whicli cozidrictiori takes place by mear~s of lioles atid electroris), and ari: irlsLilatcar (meaning a rxiaÃerial that is a poor conductor of 1~eat energy and electric.ity). They were designed as rnitaiat.Ãrre Faraday ca.<weS in order to facilitate detect:ian in :~::tR:l. The particles shield (r~~eariirig protect; screexi; block, absorb, avoid, or Ã?themise prevent the e1fec.tS of) tlie oscillating niagnet7c fields in MNM that arise fi"ori-r radio frequcricy (l~F) ~~a~lses aÃid t~~~.~wt~.et:ic ~iclcl ;.~r~.die.i~ts: in an itz~~.~~ir~.
; sequence. rl"~l~is sl~ieldirx4~;
occurs as a result of eddy ctgrretZts (meaning circulating currents induced izl a coiiductor aiioved tlia-nugh a ma>r~etic tÃelcl, or ~~l~icii is subjected to a war-virig ma#.~rietic field) generated in the -l:raÃiie cyf tE-ie partic.lc that induce a local magnetic field; which interferes dest.nic:t:ivcly witli the exxerrral mag.rret:ic 1=ie1d.
100601 In one aspect, the present invention describes the self-assembly of 3D
metallic particles frorn 2D ~.~hoÃo[ithogra:pli ically or eleetrtilithogral.~hica.lly micropatterned precLrrsors. The terms "ph~~to1ithography", .:pbotÃa-1ithography", ar "photolithographic process" refer to a. lit~ographic techiiique in which precise piatterns are created ori, substrates, sucb as mctaIs or rcsiris, through the use of photograpbically6 prnducecl masks. Typically, a SÃrbstrate is coated with a photoresist tri.m, wliich is dn'ed or hardened, and theti exposed through irradiation by light, strch as ultraviolet light, shiriiaig thruÃrgh tl-ic photomask.Thc urYprotecrecl areas then arÃ;
removed, tastrally through ctching, which leaves tlic desired pat:terriS. Electror~
beam lithography may also be used to create the per-f'arat:ions or pores.
10051] Tl7c. particles of tlic prcScrit invention are. Self-ti}lding and self-assembling.
The at least one hinge of these Stnactr:rres coriiprises a ma.terÃa.l, inclLr+:lir~g btit not limited to, a solder (meaning an a1lo~= formulated to have a specific melting point for u:+c J17 ]C1n1t7E_~', i'Ãletal;;), a 11#;tall9c alloy (mcs311Ã3g a Ã311)S.tui'e ['o31t'c713iÃ3.~w, twC? or 31C]'e nieiall:ic eieriÃcrÃts or gÃ-icta.llic aid nc3rlrneta.ii:ic clenicnts usually liÃsed together or di4so1v-in~,r irÃto each otkÃer .wlieti molten), apc~lymer or a. glass thai cai be tiqr.Ãetyed.
The surfiac:.e tension nl" the liquid hinge provides the force necessary to fold tlle "D
template iritn the 3 I) particles.
1.0062:1 In another aspect, after se(faassemti1y, the fiilable ceÃiter chamber of the particles of tiie present iiveitiori is available as a vessel for encapsulation caf t(-ic.rapeutic agents. As tised E-iereiii; the ÃerrYi 'the.ra.peutic agent"
refers, to any pharma~cuÃical agent, composition, gene, prot.caÃi, cell, molecule, or subst.ance that can be Lised to treat, control or preverxt a disease, xiiedica.l cc?xditioxi or disorder. The term "cornpnsitit:Ãn" refers to a rnia.Ãu.re of arÃgredients. The t:erri-i "
pharrriaceutic.at composition," as Lasc:d hereiri; reters to a c.oinposition, which has under gone federal aegulatory= review, "I'lie t.errii "treat" or "treating" iicltÃr;ies abrogating, substantially inhibitin~;, slowing or reversing the ~.~~c~t~essinrt of.' a coriditiori 5ubsÃantially arÃieI:ioratino clirÃic:al or syniptoa~is of a condition, and subyt:arÃtia.lly pr-eventirxgthe appearance of clinical or symptt~nis of a c:c}adition. The amount of a therapeutic agent that resÃ.rl.i in a thz.rapeutic or beneficial efTect following its admiriÃstration to a aub~jc ct, iÃicludirig humans, is a `'therapetirÃic amoÃ.ant" or "pharmaceutically effective arnouÃ1t".
The thera~etrÃic or beÃ~eficial etl'ect can be c.a.rriÃxg, rxiiriimizrrig, preventing ar ameliorating a disease or tlisordcr, or may have any. other iherapOrÃtiC or pharÃnac.c>trtical beneficial e6c;t,t. The terÃi "disease" tiÃ
">dis~.~Ãdr:*.Ã=," as r~sed herein, refers to ai impairment of healtli or a condition of abnormal fr.nct:icming.
"I'}ie term :.syrr.drorne," as ti:~~~i herein, ret'ers to a patternof symptoms indicative of some disease or c;ondit'itgn. The term "injur-v," as usc ki hereiii. refers to d~-nage or barrÃi to :-l sÃmeturc or tianction of the body calÃsed by an outside agerit or force, which may be physical or chemical. The temi "condition," as a.rsed herein, refers to avar-ieq= of health states and is meant to include disorders, diseases, or injuries ca.usCd by any underlying mechanism or tfisordcr, arict includes the prcir~iotiori of h"l thy tissues and cargaÃis.
100631 In so.me era-ibod.irnents, the fillabie center cl3arribcr of the particles can be i-ascd to encapsulate sLaclz therapeutic ag;ents as _pliarr-fraceratical agents or drugs, liairx4;
iis4ue. 4rels and polymers, tvliicli subsequently are rc:ieased in sitr.i. As used liereiri,, the t.er.an "pnlyrncr4J refers to aÃ-iatri.r-al. or- syr-iÃhciic compound consisting of long, repeated and somcti:mcs branched chains, built rip from small suburlits c.a.iled morlor7lcr.s.
Nattrr-al polymers incla.ide proteiris (polymer of a:riiiiio acids) &
ceI(Lilose (polymer of s~igar riiolecules}. I'here are mariy cxariiples of synthetic po(ymcrs, f00641In so.rnc embodimcrits, #i:znctional'==: cells +(c.~~ ;panc.rcatic is.let cells, ncurcanal :l't 122 cells) can be ericapsuiated t~~r in vib-o and in vivo release witli or without immunosuppa-ession. Sr.wh par-ticles cati be administered to a subject in xieed tl-rereofi by iiiic,roir~jcction, eithcr as a siÃ-igle biocoritairicr or as :1. ;rou~~
of"biocontairlers aÃid are useful for imaging, diagnostics-, and t:hera.peutics.
100651 For exaniple, in oae embodimerit, f1ie iritericars of a multitude ~f particles were filled with cells that were cziibedded in agcl. I'hese cells could be released by imrrie.a=sing the biocontainer in ari ap}irc3pfiatc so(verat. `I'he rYiagrictic resoi-ratic,c (MR) images of the pailiclcs embedded in tlr.lidic media srr~gest RF shielding and a sr.e4ceptibility effect, prcg-v-iding tharac.tera ;tic 1-aypointc:nsitv (+:larknc ss) wiihin tbc particle, thereby alk-nving the particles to be easi ly detccted, 'I'lii-, dcmoÃistraticrl is the first st:ep toward the design ~~f "ID, micr~~laatterncd, rioriairiva.sively trackable, encapsulation and delivery deyricca, 100661 fi'he present invcntiozi provides a three-dimensional particle i:omprising a plurality ot ovo-dimensional 1`a.ces capable of self-folding to form a holimv Ãnteriar, wherein asi~e of the particle is mic.rosc.a(e or nanc}sc.ale. The particle preferably ranges in size from 1 nna to 2 mm.
100671 The part:iclc flrrtltcr comprises at lea-st oric hinge, whÃcli may be ccrriipriscd of any liquifiable mat.erial. For example, the hirage mav be apolvriier, a gel, glass ar a m etal .
1Ã1068_1 The parÃicle of the present inverit:iori Iias any shape, but prefirablv has ster:fac.es fc?r'ming apolyhtadra( shape, sr.rch as a crrbc. The particle's two-dimensional iaces are pat_temed with perf1wations oi` pores. These:: perforations or pores may be created phntolÃt:howrapl-iÃcally, electroiithog;raphÃcally or using electron beam lithography. These perforations or pores 11az~e a size ranging frorn 1bOÃJt 0.1 tirl-l tÃa about 1crn. :('ret:erably, these perforationS or pores have a size frori-i about 1.0 rti.m to abaui :l cm.
1.0069:1 '1'he liai-ticte (container) of the preseiit invention may be fabricated from aiiir trtaÃerial, laut preferably at least c~iie material selected 1=roiti the group consisting caf a i-zietal, a polymer, a glass, a SemicondÃtci:or, an ii-isÃalator, and co.mbinations ttiereof.
'I'lie paa-ticle n~ay also comprise active electronic or sernicondiÃctor ecyiiipcyrietits suc1i as transistors, sensors: aetuator:=, lig}ii emitting, diodes, photodiÃ?des and solar cells. If the particle is metal, such metal iiiay [~e copper oà rric_h..el. In otie embodiment, t~~e particle is a. Faraday cage. In another einbodi.itient; the parti~~e a~av !~e coated with a bioeompatible material, such as a metal, a pol~iner, or a Ã:,onibination thereof The panic1~ ~iia~~ further be associated wil(i abiowrtsor.
1[)070] '~(`he particle may#iirtl-acr comprise at least oÃ~~ substance, sue~l as a therapeutic agent, eiicapsulated within the paÃticis;.. The therapeutic agent nlay be a cell, zt chemical or biological a~~~.ait, a pharmaceutical a.gent, a c:oml?ositi(iri, a tisstÃ.e, a gel, aiid a polymer. In certaiii embodimerits, perforations or pores in the two-dimensional faces of the particle allow release of the contents of-the particle.
10071.1 The particle of the present invention may, be adminis"tered to a subject. 111 such ~~~i etnbodiiiient, the loca.tio~i of the pw-ticle in the subject may be non.ainvasively tracked bv magnetic resoÃiaÃice imag-ing or CAI, scar~ (Crl"). 'l`~e particle may be imaged with iieS.sat:ive contrast relative to back<wr~~~~nd or positive contrast relative to ba[:kgÃ"Z713t7d.
100721 liz aiiotlier ~rnbcdiment of the invention, the particle additioÃially con1~.~rises a radio frequency tag, wherein ttie sa.ibstata~~e may be released upoÃ~ the particle's expoSÃirÃ: to a pro-Selected frequency.
100731 In a t:r.rr't~~~r embodi.F~ient of the partic:le Ã.xf the rnvel-1t.i011, the substance rr~ay be released Lipnri the particle's exposure to electrori-ia(Si-ietic radiation, wI-rich r~iav be tri~.rger ed r-etrioiely. The eleetrcsr~~agcic:iic radiation i-a7av r=arigc from lK I-iz to 1Peta Uiz 1U074J In a:fiÃrrÃber cgnboclirnent of the particle of the .invent-iorl, the srrbstarice may be released tilaort the particle's exposure to indLactive 1~eat:ir~~~;_ Sueli iriductive heating niav be taig*..~ereci reriiotely.
100751 '~('l~e preserzt irivcritioÃi also provides a incttlod. o-t.' fabricating a tklree_ dinier7sional partic[c cÃ3rzilarising a Ãiiiilt:itLide of Ã~.vowdiinensional faces that )`or-r-fi a hollow l,olyhedaal sliape a:rid contairiirig a tillable center chantber. This txiethod cor-aprises, the steps, (a) fabricating a multitude of two dimensional faces;
(Y) patterning the fabricated t~~~Ã~-dir~ieri~:ir~i~.al #ac.es; {~.) paÃt:er-riiri~.~~ at least Ã~rie I-rirt~~e ~ari tfic patterned two dimensional fiace to form a hiÃi4>ed edge> (d) joinint; a hir~~,~ed edge of a first pattemrrd two dimensioÃia.1 face to a birx(;ed edge of a secand patterried two diriierisioiia( t:ace to torrii a kiirigedjoizit, (e) r~~~~eatirig step (d) to fori~i a tNvo dimensior7al precursor teÃiiplate 1~avirig hing;ed joii-iÃs bctweeri adjjacent t,~~Fo dimensional faces; and (f) lic:ir.rel=='ying the hinges of the t.~vc>-dimertsion;rl tr~niplate usint;
heat to iiiiiizite selrr#'-fulding. This method a.(1~)~,vs the pal tic.leio self-assemble.
100761 In one, embodiment of this niethod.t the hinges of step (e) comprise a mateCiaJ that can be liqa.Ãetied. The mat.ctial rnay be a solder, a inetallic a.llo`, a polyn:rer or a glass.
10077] In mothcr e.m$.~odimerit of this motl=aod, step (a.) ftirther comprises tl rc. ste.ps:
(i ) spimiing a sacrificial film ori a substrate to form a first (aver; (Ã.i) layering a conductive second layer cgri the first iayen, and (iii) patterning the layered substrate by phoCo1 i ihtagr;:tphy.
10078] liz tlicse met1iods, thc particle has a size that is riiicroscale or rraÃlowcaIe and may have two-dimensional faces patt.cnacd Nv it.la perf~~arations or pores, whictt may be created p}totolithcggraphically, and may vary in size from about 0. 1 nrn. to about 100 niic.roiis. The particles of tliese r~ietliods may be a Faraday cagÃ;.
100791 '~1'1Ãe invention further cumpz.ises a m~tht)d of irÃ-iagiÃig a.
particle of the iÃiveÃa:iorà that }ias beet-i igi-iplaÃited it-iÃeà a Sr:abiect coÃ~ipi-ising;
the steps of: (i) Irladirig the hollow irÃterior of the particle -witlà at least one substance tE3 form a loaded part7cle; (it) adrzÃ.ir7:ist.ering the loaded par-ticle to the su~ject; and (iii) rionrcinvasively tracking the particle of Ste}i (ii) in the SubjJect $?v magnetic t'esoriariee :irnag-ing.
1n one ernbodirlient, the particle has pertoratiotiS or pores in its two-dimeiisioria.l faces that allow release of the sulasÃance W tfie hollow iriterÃor. In orie eÃiibodiÃiierit: at least oÃ-re substance of st~-p (i ):is a therapeutic agerii. 1'he therapeutic ;Ã.oerat may be a cell, a pharmaceutical agerÃt., a coÃtiposiÃion, a tiqsue; age[, arid a p~~fymer.
~00S0:1 The meti-aocls of the invention also comprise a met}Ãod of treating a condition co181p1"ÃSltl%M introducing 9.ilto aii a.t1.tI71al til I1eed of tl"ea:l'rtlellt at least. C)ile particle of the .iriveritioti encapsulaÃ:iracy :Ã. 001Ã1posit:io11, wlier-eitr t1le composition iS
released through orie or more pores witlain the par-t:icie into the inammal in an amount sufficient tc) treat tlhe coridition. 'I'l~e pharmaceutical coÃ-zipÃaSitÃoÃi rii~~~ be contained within one or Ã-iiore m:icg-e~~~eads. In one embodiment ot'thi~ inethod, the coridition is diabetes, ar7d the composition is one or more irlsr.llin-Sec:rctirr~ cells.
1Ã}041.] The invenfioÃ-i further provides a method for imq ~ing a ~,~tii;1 of f the iÃiveÃiÃion that has been introduced into a ma.nimal cor-ri~.~rises using ma~,~netic resonance ima-ging.
10082.1 The iriver-itivr~ further provides a method for= rargeting the particle of claim 1 to a. cell within a sLÃ~jec t eomprisirig the steps of: a) a.ttaching t(i the particle an antibody aga:inst aii atitigeÃ~ specific to t(ie cell; and b) introducing the particle into the mammal, whereizi the particle is targeted to the cell, 10083_1 1n aÃiother aspect, cells withiii or proximal to implanted putsc:le;
of the present invention can be irnaged by MRI to evaltaate tlÃC efficacy of the implant and the condition of the encapstÃlated cells, 10084] The invention also provides a ii-acthocl of dc.Iiver-ing orte or more particles of the invention to a suL~je:et, wIie;reirà the particle is pro<Trami ~cd to remotely rel e.-Ãse, one or more reagents at a.n~= specitit, time and at any specific spatial location.
(n. one Ã:rribodinienà of this niethot{, the particle is i`eiiaotely guided a~id Una{,~ed tisir~g 1A.RI car C"1`.
100851 Also pr~oviclecl is a method czf'reieasirtg a Ã:.ontrast~~,',ent from the pi.rticlc: of t(-ic iriventioti or of pg-Ã~vidisig contrast to allow ?,~I:IU or C,.l' iniat;int; of its coÃiterits or of stabstattces`Niit_[iin its vicitiiÃy_ l.00S6:1 A metliod is also provided for conductir~Ig non-invasive biopsy c?f znic;rosurgery, coiiiprisiÃig dÃrectin~; the particles to a site within a si:il~jec-Ã using reÃiiÃitc r~~eaÃ7S, allowing the particle to captiire i~i-ic or ÃTloÃ-c stabstances i=re~i-ii itle site, aild obtaining ttie substance from the part:icle.
1.0057:1 Where a range of values is provided, it is iixiderstÃ?od that eacli intervening v<ilue, to ttie tcÃit}i of the tjÃiit cyl' ÃbC lower IiÃviit Ã.MlesS the context clearly dictates other.:vise, between the ~ipper and lo-~vcr fa~iiit of that ra.rig~ and any ot~~er stated or iÃiten,cning value in that stated range is encompassed witliin the invention.
Tlie ÃrF~-per atid 1ower liiiiit: of these SÃ~ialler raiiges, wlaich may iiidep~iideiit:ly be included iri ttie snialler rariges, is also encompassed wit&i:iÃa the :iÃivent:ion, subject to ariy specifically excluded limit in the stated range. Where the stated range includes orte or both of the liniits; ratiges Ã;\c:luding either both of tliose iiic.luded Iiinits arÃ;
also incltid.ed in the i ~iveiiÃion.
10088] Uii1ess defined othet-w-ise, all technical and scieÃitifte ternis Lised hereiii have the same meaning as comtiioÃaly understood by. one of ordinall= skill in the art to whic:li this inventioi} belongs. Although any methods anci materials similar or ecluivalerit to tliose described fierciii can als~~ be LÃscel in the practice or testing of tIle preseiit inventiozi, ttie preferred methods and materials are now deSc:rÃbed.
All pUb(ic;atior~s mentioa-ied herein are incorporated hcrÃ;ir~ by reference to disclose ~i-ici described the met1iods andiÃ.~r matci'ia[s in connection witli wliic.li thc pÃÃblicatialis are clt:ed.
10089] lt must be noted that a.s Lased, herein and in the appended c:lairris, t.llc singular forms .<a', .zagid"> and "tlae" inc(ucle p(ara1 referearc.e;; Linlesw t1ic context clearIv dictates othe~~,vise. All techniual cmd sciei-it.ific tc::ri7is Ãi.:aed herein have the sarne Ãrl eaninu;.
100901 The publications discussed het-ei~~ are provided solely for theÃr discloswre piior to the filing date of the present applacataoÃa. Notfiii-i4,, here:i11 is to be construed as an aclmiss:iozi that the p:res~rià :iÃiveÃitioii is iiot entitled to antedate such publication by virtue of prior invention. Fua~ther; the dates of publication provided inay be different froti7 the actual priblicatioÃi dates which may need to be irideperideritly corllirmed.
EXAMPLES
10091_1 The follo'wing c;xaniples are ptit forth to provide those of ordinary skill in t:tie art avÃth a complete disclosLzt'e and d~script:ioii of ~~ow to make and Lise ttie preseiit i~~~~enÃion, ~iid are Ãiot iiiteÃided to liniit the scope ol:'what. the inventors regard as their i~ive tition nor are they, intoÃided to repre;;ent that the experiments bÃ;1aw are all or t1le only exl:rerimcnts pcrtormeci. Efforts I~ave been made to ensiir~ accuracy with respect to numbers used (e.g. amz ~a.Ãnts, temperature, etc.) but some experimental errors and ~~eviat:ions :~hoL~ld be accounted for, Unless indicated otherwise, parts are parts by weight, ancs(Ã;cula.r weight is weight average molecular weight, temperature is in degrees Centigrade, and pressLwe is at or r~ear atiiiospheric.
E-varirpk- I. Tabric:irtio~n r,+af.thc C.:orataM~~~s (Particles) 1.0092.1 Fi ;uae i is a schematic dia~;;raan of the process flow used to fabricate tlle ~ C3 containers of the present inyrt.ntic3t7.1110931 First, a 5pAm thick sacrificial laver of poIymetlivl methaerylaÃc. (1'MNtA, MW=996 K) Nvas speiÃi on a silicon substrate. The term "spinning" as i.ised liereil) refers to a process whereby a fltÃid is dropped on a rotating substratc. A '1 ? in-n laver of chrtarnium (Cr) and a 100 nii-i thick layer of copper Wt~) were evaporated on top of the PMMA coated wafer. 'rhe ('r layer functions as an adhesive promoter while tile Cu layer fiiiictiotis as a cotiductive seed laver for subsequent electrodeposition. Since it is necessary to etch tl-ic Cr and Cti later in t.lic process, it is nec:.-es&iry to minimize t}icli` thickness to achieve a i`apid etÃ;(a.. l-Iowea=er, c) minimize the electrical resistance of the film across the wafer dLrrÃ.raig eleeÃ.rodepositÃon, tlae rzaaterial itaickraess has to be increased. A thickness of 125 tir-ii was cieei-a7erl optitnal fior the prewnt application.
After t}iiri film Ãiepositiora, the sLibstr-ate was patter-raed using pI-rotofiihÃ?gral,lay. 'f .he photoresist Shipley SPR220 (Rolarri and Haas, www.rolarialaias.coni;l was first spun (ara the wafer substrate, the thickness ~~f the p(aotcaresist was ccaratrolled by changing the spiti. speed aiid the titarzaber of coats. After a soft bake, the resist was exposed to tJV
light using a aiaask: alig.raer. `I'1ae photo.mask used to pat-terta the resisà was a transparency mask witli six 200 Ãarxt squares spaced 20 pm apart. After exposure, the wafer was developed and tlic thickness ol'the resist was xneasuredusing an Alpba-Step p.r=rs#ilomete.a=. 'I'lieai; e[ectrc3deposit7c3ri was used to biai_ld pattern ttae metallic faces of tfic container in the photoresist Ãiiofd up to a taeio;ht oi' 7r15 p.m, usir~~v; eortlftxerci~l elect:rolytic solut:ioais ("I~eclanic: Iaxc: Nvww.teclanic,corai) containing the metal ions of c(ioic;e. (:`u was electrodeposited -followed by a -kliaii layer (about l. pm) of gold (Au) to fo.a=ri7 r7oaa-~~iagaactic coraiait-iers at-id a tliiÃi laver- (abOr.It 1 pi-ail) of nicJ<el (Ni) to fabricate magnetic cc4r7tainers. The Atr was Lased, to protect the CLr sL7r~acr~ from subsequent etr;liiiag steps witl r:Ã;riÃlÃ;r it itiert.
J0it941 A secorid round of ~.,hoÃo[ithogr-a.pl~~~ Nvas performed in order to pattern the hinges. A second layer of ;ST'.R200 was spLira on t.lae substrate and a hinge photonlask was rÃsed to pattern the hinges. The hinge mask consisted of two 1;Ãnwis of'hÃr~ges (50 x 160 (a.tni"aÃaÃl 25 x 160 pna). The wider hsnges were at the interfaces of ad~lacerat fa.cez w l~ile the narrower }a~r~-es were at the ed~;es of tlie taces. AligrarneÃrt marks were used to ensure perfect aligrimerit of tlie 1~ingles to the faces of the 2D
pr~ectrrsor=. Prior to hiÃagÃ,~ elcctrociepoiitiorr, the exposed t_~`tr arrcl Cr in the area of the hlÃages were e c:heci rasiiag commercial etclaarats (APS-100 for Cu arad CRE-473 for Cr, 'I'ec}ariic, IÃie>
~~~~~iv.teÃ:.hnic.Ã:..Ã-araa). Alt[ioLagla tlae etchants 1iave a high selectivity of Cu or Cr ~~,ith respect to Ni or Atr, the etch tlri-ac was c}ptimizc.d, to mirrirniz~ ~~~~~age to the Ni or Cu/A.u frame of the c:~~~taiiier. Pa,tre; tlnl.m.p. 232 -`Qor tira/(Ã;ar.l 1Sn/Pb: na,t.~. 183 'C3 solder was then electroplated in the hinge regioris. The lreiglit of the hinges was approxiiiiate[y 5Lim to about. I -5 pr.a-i depending on tlic face pattern aDd ilie type cYl metal Laaeci (wettinM, or iioÃircweÃting). .A_#:ter electa=odep sition, the origirial seed laver was etched ~iid the 2D precursor ten7plaie was itt~tnersed iri asolution of N-Methyl Py.a=.a=ol:icio.aic (N-MI', which dissc~~~~es, the sacrificial PMMA la;:erw) to release the precursors fro:rtti the wafer. Approximately 50 precursors then were scattered in a sinal( ca-vstallizin4,~ disli LasiÃig a pipette. A very ii~in laver of RMA-2 flux, (lzidiLim Corporation, www.irzdium.Ã:,oÃrt, used tt) dissolve any oxide it7rttted c~ii the solder) lvas p(yured into tE-ie diSli. The dish was theÃi heated to 100 'C t'or abotit ?Ãrliri to about 3 mfn and tlieÃi rainped rgp tc) about `.2'50 'C t~~ about-300 'C for 20 SeÃ:,onds. BeeaÃise of the low voltiÃaie of fltix, the agitaticin, was sufficxettt to correct for defects in the fiolding but iiot large eriotigb to cat~se, the crosses to ct)(lide iÃit:e~ each ot:lier atid becoi~~~ fi-iserl.
The n7olÃen solder generated ttie l=o.i-ce needed to fold the 2D l,recur~ors irito 3D
containers. 011 Ã:..ooliiig: ttie cotataiÃiers were perÃ~anciitly }ield to(;et.her by solid solder rti iiges.
1[)095] Dia.magnetic copper (Cu) containers were fab.ricated witli linear dii-iieÃiSioÃis of about 200 pli-a (rvhcrL one picometer isff'M ii-aeterj. As compared to smaller or lar.p,er sized biocapsules, the 200 pm size provides tlic maxi~~~~~~
crÃcapsula.tion volumc while still allowiÃig the ditTtiSiott of oxy<deÃ~ and nutrients to the cells.
It is kt~ow-11 that if cells are more than about 150 liÃri to a1~oLit 2100 ~tm away frortl the nearest blood vessel, the environment becomes hypoxic rR.H. Thc3n-iliiison wid L:.R Gray~
Brit. J.
C'anc:c;r. Dec. 9, 539 ( 1t355)i. In princip(e, the fabrication strategy desci ibed hereiii also would work oÃi. smaller or larg,-er size scales in. the design of containerS
for otIier a.p}:slications. The 1Ãnea:r diitaenGioii of the container was orderS of magnitude t~-naller than the wavelength of the oscillating ~~agnÃ;tic field at 500 MH-z, which is the hÃ~~heSt operating frequency in our magnetic rosoriag7cc (ANI-R) scariiiers, Het1ce:
ttie size of the perforatiotis oti ttie faces of ttie container }iacl no cietrinientaI effect on the shielding characteristics of the container. The thickness of flie ~aces of the container was clc;.signc;d to be larger thati the conductor skiri de:ptli at the frÃ;quÃ;ncy ot'th<; racliation.
T}ie term "skin depth" refers to a measure of tlie average depth of penetratic~i-i of an c.lec.tÃ'omtzgnc:tic: ti.eld.irato a rnat.eÃ-i.a(. lt. is defined as i.he depth at whic;li the priniary electromagnetic (EM) t:Ãeld is attenuated byrclcc,re:lses to (lle) of the field at the sÃar#"ace, or to appi-rxxi~~iatciy 37 f.~ of it, -vaiue at t:i-ie scÃrf"a.cc of the shield (A.
'1'sai.iovtch; I:Iectromatgltefic JVtr=e1esa .,41)1.Ãlicctfions (Muwer Acatlertiiic Publishers, M.A, I ~99"),)_ A tl7:icker container also has lov~er conductor resistance, ensa.ÃrÃng t(iat the eddy currents persist long enough to maintain shielding during the time of image acclttisiÃzoÃi, I'lic sk.iÃ~ ~eptli of Cu at 500 M:E~-lz is about 2.9 ptt-i (C. Kittel, Intr(xlaretton (Wiley, New York, ed,, at 7(1995 )): }ieÃtce, containers were designed to ~~ve, frames with thickness ranging from about 7 pÃnto about 1.5 pm.
1[)096] :[terrornagnetic Ã-iickel (N:i} containers in aclditioti to the cliarnagneÃic t'l-Ã
containers described above were fabricated to investigate tlic effect of mag.ti~
susceptibility ori the MR images of the contaiÃaer. Map-netic field distcÃrtions laut nat IiÃiiited to, shape, aiiiplit~ide az~d phase clisioÃ-iiotis, resulting from the cliffererÃces in inagtÃetic susceptibility betvkeen an ob~-jeci and its surrouncling Ã-iiec~ium cause a loss of phase coherence in the magnetization of the sample. Since the magnetic susceptibility ot'Cti is comparable to that of -~,vateÃ~, while tliat of Ni is orders of mallnitude higher than that of waÃeÃ-, a more pron~.~Ãunced distcartitÃÃ1 was expected for Ni containers in ~queoLas media (L.W. Bartels, et aI., J. l EUc. lntei-v=
licOiol. 1 ~~: _3 )65 (2001)).
100971 fi'he strateg~., tise.d to fabricate both the. Cri and. Ni containers ià Ãvralved the auto-folcling, of 2L) metallic precursors using capillary forces. "Capillary ac:tion''', capillarity ' or c:apÃllaR, motion, wliicll are used interc~han1geably lierein to refer to the abilitv of a narrow ttzl,~e to draw a liquid upwards ~.~.~ainst the force of gravity, occurs when the adhesivc iÃitcmiolecular forces between tEic ligL-id aricl a solid are stralt.;cr t1iaai the cohesive interrnolecula:r forces within the liquid. '-['he same effect is what cauSr S porous materials to 5oak. L7p liclt7icls. Previous demonstrations of acÃto-lolding include the actuation of rÃiicroziicter size t.ompoÃaeÃats and the assembly of ~}D c~niplex stnÃc:turtas (E. Smtala,et al, S'r:ieÃrce 268: 1735 0995)7 P.W. Brec.ti. et al., J.
<~~r~:rcr- lc~~~tr~r3rrrt~~:lr. .st 4: 170 (I.~1~~:~) K.F. Harsh ~.t~ :;~l.R
At'ftÃ~tfÃ~f.>~~ .4 ~: '?:>7 (1 E399)l;_.. [_:. Htii et a1., If;t~.fsl .a' ~`} IÃrt. [.YrartjÃ; On X-fict'o .f.kc:ir'c~ Nfec:hcrÃfiec:zl t~.xttts, 602 (2000), D. I=4 (Ft-aeias, ei: al Adv. A-iÃrler . -l 4 ; 23 '? (2002)).
[Ut~98J .'4ccni-dirig to oiic aspect of the preseiii i~~~~erition, _>D. I-ioIIow, peÃ-)`ora[ed containers were fabricated from ~.D precursors. 'riie process used to fabricate the 243 precursors, which is an extension of the process described in Exa.inple 1, and reqÃiired severzil. additive lavcrs, two photolithex;ra.py steps, tNvo electzodepositikiti steps, and a.
precise sequence of subiracÃ.ive processes. Briefly, the process i~ivo_lved i3ati:criiirig the metallic 27D faces rflsi~~g pbotolithography and electrodeposition on top of a sacri.ficiai Iaver. The versatiiitv of the strateg y was demonstrated bv fabricating precursors wNinse faces c,orrt:aiiied two different patterÃis-----c~~~e pattern comprised a square ti-ar~~e with open faces, while t}ie otl~er consisted of a m:icroscale cross shaped patterii iti the center of each face. In asecoiid laver of pIiotoresist, hinges patterned on the edges of the frames. `I'Iie width oi'the lai~ige between t-~~o a.djacezit. -t.'aces wa.s t~-vice ttie widÃlr of the Niinge at tl-ie edges so tl-iat aCl Iiirigcci ,joia-iÃs I-iad equal solder volume i:~pw~
foidin9: the solder vo1~ime was critical to eaast7re a folding angle of 90' (R.R.A. Syms, et al...r .I. I'?; 387 (2003)). After the 1iÃziges were t.,a.tterried, the 213 precttrsors were lifted cff tt-ic wafer by dissoltitiori of the sactiticial layer, '1`he containers avere sel.tmassembled by 1~eating the precursors abc~ve, the rnelting point of the solder, wherein ti-ic liqa,tid solder with bigh surface teiision generated t1l.e :i=or-oe rc:c;tiired to f-o(ci ad~ja-cc:nt faces of the precursor.
1.00991 Fi,,g. 2A sbows aii optical ima<we of a c.oilect.icari of containers that were fabricated tising tt7e process outlined above. The fabrication ttrategy allows a large trUmb~,'r of containers to be constructed in a single process riin. The primary yield-linziting factor was the error in estiiiiatirig tiic voltitiic of the solder to be electr~~~eposited at eacb iiiiige. The spaciiag between t~~e adjacent taces was also critical-ysrhen the gap between faces was either too I'''.argx or when the faces were ft~sed, the yield of folding was greatly $iniited. Fig. 2B=22D show optical and SEM
imagges ot'the mit;respattemed containers at different stages of the fabricatÃon process:
the 21) pr-ecÃir5oi- with e1eÃ;trodeposited fiiceti, ilic preÃ;urtior witli i.~tc:es arid h:ir~geS, aTici t (-ie folded container.
1001001 Although a~~ opeti-Caced c.oa7taiticr B, iaot ideal for= ari enc.ipsuilation device, since it is corisiderablyleaky, opcn-1'aced containers were filled for easy visualizatiO11 of their cont:entS. For M vivo applicatioris it .rtiay be de5irabIe to use the described strategy to construct containers wiili selectivelv sea(ed or micro/nano perfcyrated faces, and :(=a:1?ricate r~iore c.oztiplcti; polylicdral containers with r-oLi.rtded vertices. A.11 open-faced container (Fig. 3A) was loaded ~Nvith rnic.r=o"bea.ds since niany cellular dcliaeo., techniques use microbeads witti cells adhered to t.faeir surtace. In order to load the corrtainer with microbeads, a susperision of tl-w beads irt ethatio1 ,.va=
pipetted orito the car7iaitie.r. '1':1-le suspension entered tl-ie container as a r-csult of capill~~~~ forces. When tft.c ethanol evaporated, the beads were lield together by weak van der Waals forces 4at~e~~ir~~ the weak intermolecular Rorces that arise frorxi the t:ra:risieiit polarization of a giveit. rtiolcctile ixito a dipole) (Fig. 3:B); the glass beads cnÃ.rid (ae released by agptatiorl of t Nic cor7 ta'a tie.r.
1.001011 In order to dcnionstrate cellular encapsulation, MD.~ A,1.13-23t breast cancer cel:ls in an extracull.trlar= matrix (EC"N't) suspÃ;i-asiori at 4 'C, were ~~~~~ied in t1w contairiers (Fisn). 3Q, As tzSed 1-rcreitt, the terrii extracelltt[ar rt~atrix refers to the complex staÃrÃ:.tÃrra1 entity sÃÃrroi-rrid~~ig aaid, sÃrpportirig cells that a:re touzid wÃtliiri mamma1ian, tissues, as well as one or more of its constituciits incltrwiÃng, bÃ7t not IÃmiteci to, collagen, elastin, fibronectin arid Iwninin. MDA-4\4Ba22s1 cells are representative of rapidl'y proliferating cells and irximortalizcd cells, sLrch as P'I'O
cells, used in diabetes therapy, and stem cells used M rerwenerratiorr. On incubation at 37 'C for 5 miai, the ECM sÃrStrenaic3n gellÃ;d: the cells were retained in the biocontainer= and c:or.rld be released by immersing tttc corita-incr in wairii cell ct.rltÃirc mcdiÃrrn (Fig. 3D), It was also possible to load the biocontainers witli a celI-ln,C:N-1 suspension withiÃi an agarc?s~
~avity. lrr this casc:, a suspensiori of 5% agarose gel was tirst micropipetted ('60 prn tip) into the c:ontrziticr trsi~g a stcrcotac:tic: manipulator. The gel adhered ta,~ the sides tif t}ie container thereby 5ea(ing the faces ancl Icai>iÃi~,= a void in the center of t1x container. 'J`hc cell-l:C"M strsperisiozt was t.lrt:.ra rngcroirrjectcd into this void, w}ricli w-as then sealed with a microdrop of awY.t.roSe M~~~ci.
1001021 To demonsrat.e that ttie c.ell, were viable in the bio-ccamaincr arld orl release, the cells ~~~>crc stained with the fluorescerYt: dyc, C'a(ceir-i :'tM
(Sigr-fia-Aldrich), which stains positively for live cells. Fig. :'IE sltmvS calcein-staÃr-ied cells witltiri ttie biocontainer and Fig. -3F s}iows release of live cclls from the container on immersÃozt in warii7 cell cÃi1ttare ÃiicdiÃarti. 'I'lte frames of the biocoriÃaiziers t-tsed iri tliis dcrnotistr-ation kiad a tkiiÃ-i gold or platiritirYi coating orr the interior faces i=br biocoÃ~~~atilailrÃy, qiric.e gold ~iid platinum are, inert or tÃnreactivc materials. Pure tilt aitd tMr`Iea.d based solders were used to fold the cotitaixrcrs. It may be necessary to use ot:l7cr solders cnntainirro; iricr-t lncta(s Sucli as silver arid 4old for errliarrced biocompatilaility. :(t: is also possible to increase ttie biocompat:ibility of tlie containers, bv coatirtg, the entire folded container witlt a layer of an inert metal (by electrodeposition) or with polytiier5 (by ianrgtersion or vapor coaÃirig).
1001031 -NnnHinvasivc dctcci:ion ol'the c.oriiainers was dcri-iotist:ratcd by cr'Ylbecldirlg the containers in 5% agarose gel aÃid imaging them with tiIRI in a 500 X1.Hz vertical btire Brttker Avance micr+,3irnas~i~~ ~ystz.m. For tho images Shm? ir Irc rc, a. s3D FLASH
ScqtrciiGC Ivith tlre cch~.~ tilne (TE) irt the range of 4-6 ms; a repetitiorl time (TR) of 50 ms, flip-angle of -10', and a spatial resolution of 25 P111 X 215 ~ÃÃn X 20-lun was rised.
The containers also were i~~~agetl using a standard spin echo wetluetice (meaning a pttlse> seclttcrice tiScci in magnetic re.soirancc irraagin<.~ based oti the detection of a st.=ain. or Hahn echo, w}iicla uses 90' radio.frequeaxcy pa.iIses to excite the ÃnagnetiSm and ozte or more 180' pt.Ã1ses to retocus the spiitS to generate signal echoes named "spin eeboeS), with sianilÃtr reaEÃ(ts. Fig. 4 shows MR images of a 900 }tm cli<t~-noter c:at.~sill~ry corrtairiirzg a CLi (Fig. 4A) and a Ni (fig. 4B) container embedded in agarose .;cl. A
characteristic signature was ol-iSen~ed for both ttie Cu a:ttd the Ni corttainers--tltere is a pronounced darkness in the region of each container. These hypointense (dar~.) signatures 1~ave bcc.i} observed before in N/1-Rl of laq4cr centimeter scale metallic coils (A. Shcnhav. H. Azhari, 3V1cr~,Fw. Rras-on. 52: 1465 (2004)). While the region of hypoiiaÃc.ÃÃsity (darkr.ae5a) in the .14IR iÃ-iizÃ~,se was coÃnpar4able -to the size of the Ãa.oTi-raaag.aaei:ic Or container, it ~,vas rnucla: larger for the feÃ=.rornzagnct:ic Ni contairaer due to a pronounced s-uscc:[Aibi1it~~ effect (L.H. Bc:ÃÃÃaett., et aI.,.l. AI)I)1.
l'h4:s. 79: 4712 (1996).
BA. Schueler, et al., .1: Xlcigrr. Resrarr, knq~,i~~~g, 9: 596 (1999)). '1 fae iÃYaages of containers :Ãraadc of a ,aveia material were similar for botli opeaa faced c.ontairaerS as well as cross taceci containers, shou;iiag t}aat the pattern of the faces had fittle bearing oza the MR si graaÃure at ttzi s size scalc.
fUO 1.041 RF shielding w.a.s simulated in a non-magnetic coaitaiÃ-ier with a {-:iaÃite eleÃ~~ent model for a 200 Sp scatc wire t`raÃxac that was excited by a linear polarized electÃ-orÃra4~netic wave. Fig. 4C-4D are simulation ro-SÃilts showing rÃiagnetic field d1st:o.a=t:innS in the vicita.itv of the cotataiÃ-ie.r :Ã.tad reduced field ma-aait-ude in the irtaÃcri(ar of the coÃlt ai t1 er.
() 1055 ] For maziv biomedical applications it is Ãaecessary toÃaon-ÃÃavasive:ly track an eÃicza.pSÃ.alatioÃi devic.e. 'I'lic (:Ã:a container of the present invention could be t-r-acl;:ed spatially aÃid teÃ-iip~.~rally with in flow thÃ=oia"tl aÃ-i S-shaped 500 }sÃ-ia diameter fluidic c.}iaÃiiicl_ The chaainel was fabricated by molding poly di~nethyl siioxane (PT~NIS) in an SU-8 pbotoresflst ÃiaoId that was patterned Ã.Ãtiing pt Ãotf-Ait1aography. The channel was seated with a. s~cc)Ãad, t'iat, oxygen plasma treated. PDNIS
la.yer, Polyethylene tubes were conaaecÃed to ttae inlet and otÃtJet ports of ttae cllaziÃie1, the channel was flEished with silicone c3i1, and the container wÃ:Ãs' inrrodiiced into the channel. Under prcwsure driveÃr flow, the container moved withira the clyanÃie1 andwaS
imaged at ditTererat positions; the secli-ieaxce of N~I:R[ images is Shcawra ira Fig. 5. This ready trackability wit~~ TNIRI. at a>erv short echo times, without the need for a contrast agent, highlig}its a ma~jor advantage of the 3D metaIlic: bioc:oniainers or the present invention as coniparcd to manly other erica.psLÃlati~.~n systems.
Exanip1e ,27, MMuMtican caf Near Alc~qtxetic H~.}l~~V in th~.~ rerg;ion r_?/`
tla~:f container:
1001061 To denaonstrate an RF `hicldirm effect, tbe near m.~gneti.c> field response in the iric..ilÃity Ã3t" t(ae coaztaiÃaeà was siztÃulat.eti usiÃa.g a fin.ite c.lcnient electroanergrÃet.ic siniultztion ptacktage, F11K{_} (EM Softwarc. & Sytitenis-SA Ltd., wwts.ic:ko.Infoi'). A
t:i-all-wavc i-iieiiiod of Ã-iioÃ-iicaits approach ~~ias used to simulate t:lae.taear mag-netic field iri t}le region of a 100 Eam wirc: fracrie wii}t wire segrnents of 8 tiiYa radius', as,'uanrng perfect electric conductors coated witli copper (coÃidÃac.tivitv ___ 5.8 13 x 10'7 `?.na`'). 'f .lae ,imulation of the cubical wire fiariie model was performed witli a linear polarized platae wave excitationat 500 ?1't~Hz; we used an excitaticati sotirce of i V/m incident oza the wire fraiia.e, with E in the z dizectioia aiid H in ttae v directioti (Fig. 4('). The copper wia-c fraria.c was assigned a relative permcability, of I. thereby yimulatiÃag only the RF shielding etTect and n~~t the susceptibility etfects. Fig. 4C shows the near magnetic fi.~eld a-espoÃ-ase in bot.ii the x-v aaid the y-z central planes.
1001071 l.aa cnnclusioÃa, the dcscrAbed sÃaate;,~~can be tased to flibric.za.ie 3D, arbitrarily trackable, biocorat~l ners that allow perfusion bctcveen the caÃatcnts of tlie biocotataiÃaer~ ~iid the si-irroÃiÃadiÃig medium.
'I'laese biocontainers are eracapstilaÃzozi devices that do iaot lose their detectability wlaezi loaded ~vitb biological coritcta.Ã. Due to their Stre.ta~;t:la and iiigii porosity; stacla m"eÃallic biocontaiiic~~~ axc: uscftÃi as basic eiet1~entS of a scatfE}ld to gt7idc the gror~~Ih of cells in _3C3. Since the fabrication strategy described. bcre is cora~patil_ale with conventional 2D
nzicrofa.bricaÃion, it also may be possible tÃ.~ add electromechanical modules for remote activation, -,vireless coraamLaiiicatioai: sigaaaJ processing, a:iad biosensing to the faces of the bicscotitai~iers,. to enable medical diag-nostics and therapeutics". The present invention also otivisioz}a that sucl~ ~'I%D cotitaiiie.rs, whicla fttracti~n as small Faraday cages, avill iiid Latility in z ~tiaer applications requiring electromagnetic s(aieldiÃig in small o>olumes.
E.~~~~~~e 3: Aficrqf4bt~ication c~~id Se?f-Avs~.anzblV qf 3d Aflcrobox~-y,v fi)r Biot~~edical AlyWcations ~r ~:1~?t=p~~t:~1 t r:~t~:
1Ã10:1.O81 The process t~sed to ttibricate the boxes consists of micrcrlabriÃ;at7on arid Sttr~ace tension driveia sclt-asscml.~Iy, [h.F. Harsh, V.M. Bright, & Y.C. I
ee. S~n.s.
Aciwricai=s A, vol. 77, 23 7-244, 1999 ; E..[.:. .[-lui, R. T. 1-;*owÃ,, & M.
S. Rodgers, in 13th .Irrt. C70.rf on pp. 602tt607; R.R.A. Syms, EM YeatÃnan, V'.M. Bi-iglii, & GM Whiiesides,,I vol.
12, pp. 387.417, ~'~(}0? 1 io fabricate atad fold a 2I3 precursor :into a 3:D
hollow ,Ãauct.ure. I"l~e fabrication process involved tfiree steps: (1) patterning the faces oti tlie 2D precursor (2) patternin(; solder hiiiges bemNreeta ttie faces, aild (3) selt=assembly of the 2D precursor (:(tig. 6) `I'be boxes self-assembled wtieii the precursors were heated above the meltiiig poiÃ-it of t}ie solder, where:in the liquid solder with high surfiace tension geiierated the force reqtgired to assemble adjacetxt sÃ.lrtaces. The fabrication strateg"y allows a large raainber of boxes to be constructed in a single process nin.
Copper (C:.`u) atid t-iic.kcl (NÃ) boxes have beeti fabricated with and without gold (Au) coated SurfiaceS (to it3crea~e baoiitettne"'a).
Defet:t Modes:
tOO1091 Several defect axiodes iverc observed (Ci4F. 7); hoo~~~ever tvhen the process was optimized yic:lÃi` as high as 90% froz~i a single wafer wer-e obt~Ã_irled.
Apa.rt f:rc.~rn obvioais defect modes Sucki as over electrodeposition that merges t:lle taccs.
;nmcnt cst'the Ia~~igcs i.vitb respect -to the f-iices, atid over or Ãs.i-ider etciiing of the sc;ed laver, the lar_gÃ;st defect gtc~ was ttae hei<:~~t of st~ld~:r`
Ã~l~:Ã;t~Ã~t~Ãi~.~~t~;;it~:d at the ~es. I:t: too Ãliticii of- too little solder is cieÃ;trodepoSited the structure OVer Or Li.rtider folds=. Ã11 order to deien-iiiiie opÃiiv~~~ira solder beaght ~'or 90"
.tiolding;. published design rules [R.R..A. Syms. C.M. Yea#.man. V.M. Briglat, & G.M. Whit:Ã.sidc.s, 3, <t~r~:rc~~~Ic~trr3r~rt ~:#rc~rric r.r1 ~~ ~., 'vol. 12, pp. :347-4:17, 200:~1 were ÃiSed. Additionally, iTi order to iiicrease the ei-ror toleraÃ-ice, liir~~~s were desi,,:ried t3etweeti adjacerit faces to be tzvice t(ic widtli of kateral solder re(;iot~s patterned along the edges of the faces.
During electrodeposition, due to the elevated temperature (200 : CY) the precursors ,rere agitated (due to convective tlow Ãn the flLaid in which the boxes were seIfwaSsembIed);
this agitation aided in correcting metastat~~e mininia (errors) aiid helped the box fold to the thermodynamic mirziÃ~iuÃii, L0atiiiw:
1001101 To demonstrate that the boxes could function as encapsulation deviÃ:es, they were loaded with a variety of medically relevant constituents inclt,tciÃ~~ gels, beads, (i.qÃaidsr and cells (Fig. 8). For easy Visiralization, boxes w-ith all opei1 tat.:..es wt;re.
tased. f=1owea=ei', .iÃa real applications, boxes witli oiily one c~~~eTt face fcir loading, w-iÃb the otkaer faces close(l or pi.grcÃLis-, wc3Ã71Ãi be Ã7;cd.
IÃ30.1.l11 The byclrogel PltarorÃic; F127 (201NO ;soltttion) e\hibits a thern-ioreversible transition from a l:iqÃa.ici solution at l~,,,v temperature (e.g. 4 "C} to an ordered micellar ctibic phase at roniÃi tetrzperattare_ This pi-operiy ra-iakes ii: vet)5 attractive in tttc storage and releast. iia datag cielgoe.ty. The 1Ãvtizoge( consisted ot':;t 20% w!"w rÃ-lixttare of1?ttaroDic.
I. l d7 l,pt:Yly(etlav1eiÃe oxide,)-blot:.k-lacaly (propylene oxide)block-poly(ethylene oxide) capolvzrÃer, (BASF C 'c~rp. -www.bas1'.t;om) in wat:er. ~l'';'tie saiaip_le was staakera tisila.4,) a vortexer to speedup the atixin.g pÃ`ocess and stored Ãtt 4 `'C be-tbÃ'e usage.
In order to load the hydrogel in the bo\, a drop of the liKltticl solution was placed sati the box. ;l7ta~
to the b;.droplailic: si~~~v~--alls t.~-1`tkie ri~~tal(ic boxq the solt.itit.~ta r-eadily eÃatered the box.
'E3t~s.a.es were also loaded witlà "~=1==;DA11!fl:B-'' 31 breast cancer cells era-ibedded ir1 exÃ.a-itcel:ltalar rnaÃ.t-.ix 41:C:1~1~ gel. (NV1:C3A-=k-1B-23 i cells a.rt-.
representative of rapidly proliferating or .iÃa-irÃ-Ãc3rtta:lired. cells such as fiTC'3 cells tased in diabetes therapy, faTid stern cells Lased in regeÃ-acrat:ioti}. FÃg;_ 8C shows: a box loaded wit:li cancer ~ell~ that were briefly suspended in ECM glel at 4 'C. 'I'lie suspextsic?tx was introduced into the lanx arÃd was kelat at 37 "C for l:5 r2aÃ.ta to allow the ECM gel to polyraa.erire. The cells were stable in ttie box, ;Ã.tid could be released (Fig. SD), by pulsatile agitation of the box. ':I'}aes~ experiments clemottst:rate that it is relatively stt-a.ightl<cyruard to load tla,e 1?o.x.~s wiÃli a variety of coristitLietiÃs.
lnterartion with RF Fiellds:
lÃ3O 1.121 Sirice tl.ae boxes ai`e rnet.aligc tl-iev iÃÃtei-at:.t witl-i :RF
fields and belifa-ve its Faraday cages. This fÃsa.ittrtr tta~ bet;n Ãtsetl Lo ctet~.~ct. Ãtrtd track.
the boxes rt.n.totelrr tÃ:;ino.
magnetic reswaztrÃce gnaaging (N'l:RI). A t}.aaÃacteristicsi<:ÃÃattare was sabserv~.acl fot bcitb the Cii and the ^+;:i boxes-there was a pronotanced darkness iTi the region of each box.
This hypointense siwt-lature t`ac.iliÃated .re-ady trackabiiity wit:li M:Ett., at siroÃ-t ecl-io tiriies., without itie need for a contrast agent, arid highlighted a major advantage of these encapsulation devices as eoÃ-irpared to existing polyme.r:ie systeais.
1001131 Since the boxes iriÃeract witlr RF fields, this firatLai-e su,= ests the possibility of inductively hea:ting the box using a remote RF field generated by passing azl aItezzrati~ii), ctirr-erit tkrre~uggh a coil [:E. J. W. `I'er Maten and I B.
M. N-Ielissen; vol. '28, nn. 2, pp. I 287-1 `?9i3, 1992; C. K. C'boLi, in -5r`!r C "orrf i=trmr=zrtrzentalroa,r trml <,WeasurfAmc?n/ Iechõ 1988; pp. 69a7 r, J S. Curran and A. N-1. Featherstone, J.. vol. 2, rio. -1: pp, 157-160, 1988; K. Hamad-4chifferlx. J.J. Schwartz, t.T Sa:zitos, S.
Zhang arid IXII. J~cobson, Ndirrric , vol. 41 5, pp. ! 321-15-5. 20021. 13oxes coiYiposed of dia.a~~~gnetic t.CLi, Au) and ferromag.Iletic (Ni) metalc were fiabri^ai:ed.
W~~~ii a box iS
placed in a coil throu,,,,,h wliicft aii AC curreat is passed, aii electromagnetic force is induced. According to Faraday and :L.:eriz-'s Law, E =-N" aIr` , (t ), where E
is eIectromotive fii}r~~~ (Ei~I:F) indueed :in the box, tp is Ã-nagrret:ic f'l~ix generated in theRF
coil. and N'is the number of the coil tums. The i'=.ndt7ced E:~~~1~' caLiscs a cLirTent to flow in the box which can cause Iie.atii-ag, TIic heat generated can be calculated a.41' = .~-~,'` R.
(2)Y. wlier'e 11 is the heating power ger~~rated by the cur-r-errts, aiid R is tlre resistance of t:1le sample.
1001141 The alternating current i-n the box is stibjec t to the skin-dept.~
~hono-nienon, i.c. tlic c::trr.re;nt den.si.tv decreases with deptli. Sirice the thickness of the surtkes of flw boxes can be controlled with a raii9c of tbicknesses limited only by the photolithographic as~ect ratio LÃsed to patt.em the 21D precursor, bka:~es may be fabricated with wall thickness con~para.b(e to ttie skiii depth to mitiimize the e1eotrical resisÃaire.c. Additionally, if the box is ferromkgg7etic (e.g, Ni}~ t.he. h eatirrg is iarcreased due to magnetic hysteresis. As the prima~y purpose of irrduction heating is to maximize the heat oncrgy generated in the box: the apert.tirc of the inductive heating coil is designed to be as small as possible wid the box needs to be fabricaied w~~~ith a matena1 that i'eattires low resistance and high pc;rnieabilitv.
1 (1{}l 151 '('wo kinds of configurations were d.eincmst.rated. [n s~iie case tl-ie btsxes were iritrntluced iiit~ a v:ial arouiid wliieli is wrappecl a e.Or~ coil th.roi.E4,~li which AC
current is passed (200 M'E4z tc) I C31-1:r, 0 i tcz I Watt). In order to itite-grate heating of t(-ic boxes wiili '?D Ixzic.rofluÃd.ics, and to maximire inductive coupling, 21:3 coils were also flabr.ieaÃed. "I'he 2I) coils ar~ fa~iicatecl photolitEio4>-rap}iically (Fi,x 9) aiid cari be made with a varieÃv of ta.irns a.iid s~acrng. 'l`he box is placed along, the central axis of the coil in order to maximize iÃiduetive heatirtg. A:liliougb the ziuÃiibez"
of turnsin the '?D coil is less tliarz tE-iat: o1'the 3D co:il, the eavÃti, of'the ?D co:il is eorriparable to the ..,ize ot-the box to maximize liiduetive coupling, hiduct:ive heatino characteristics of tlle bnNes aÃ-ld the coils are 1ieijag measured.
R.eleasiti; a chemical froni the bcax upon hÃ:atln ; ~;
1[)01161 :I.n order to demonstrate Ãhat a cliemicaJ cotald be released irnrII
t:l~e b nx-upoÃl k~eat1lig, ilie l?c~.a was loaded with a l7ydrr3gel that was dyed red.
lnyÃially, one g.a-ain (.>C l'(ti.a-ÃsiaiÃ:. F88 (Molecular weggftt.: 1. 1.400.meIting pÃ:aini.: 54 'C, obÃ_ainecl t:rom BASF) was c{issoived:ir.a 1.0 anl. of aÃ;elonc:, '('lle sainple o-vas t1~eTi heated aTid sonicated to aid disso[ui:inn. A few drops of the dye erythrosine were added to the sÃ?lutlori. Ari open fkiced box xvas loaded with. tlae dyed lavdroge1 solution usi:ti- a sy.rin~e and the box. was allowed to sit until the aÃ;etor~e evaporated. Sr~-icÃ; the hydrogel dissolves M
water, the loaded box was iÃiimersed in dÃ.~dec,iaie (hydroc'el does Ãiot dissolve i11 dodecane) and placed on a glass sd.ide Tl~~ slide was heated tÃ, 70 'C' on a hot plate and optical. photographs were taketi at 3, 7 and 10 m.ii3ÃitÃ:s. 'l`lae ge1 softened t~iid the dve was released icito the dodecane solution ffig. .10). The release of chemicals fircam other l7Nrdre~.o,els in water, as ~~~ell as c~l~tirvliri~i~; inductive RF
t~eati~~u~ of the loaded boxes, is Ã:.Lirr~iitly being ~~ivestigated. For in-vivo applicatioxis it will likely be necessary to Eiea:t tlie box approxirnateiy 10 "C' above the temperature of the human bÃ`~dy .
CONC: LU~SiONS
1001171 Tia starni-nary, a azew erac.ap sÃa.IÃttic.iia device platform that cÃ.~iaibiriÃ;s ttrc tavtarab1e aspects of three dimensionality with Si microfabrication has beerr dÃ,'rt-itansÃrafed. Development of devices rrsiÃb 7aanopt3rous faces fclr cell encapsulation therapy (without imr-ntarac3stri~press.ion) and ciesiizÃairig boxes with optinlired RF freat.ira_g profiles for reriaote relea;~~ of chemicals as in prr.3g:ress, E.Ya~~~L)le 4: R~.pnirt~.t .~ Radio-lT~rUt u~,pn. c ? Controll~.ad Nano1her Ch~-yntistr v arrd Cizeitir.c=y l I)ef.,avc~.rr, (rn .S'y bgrat~.~s, 1[){1118] Containers }rave been tab.r:icated Or.rt Of ta-leta.i, wh:icli a_lioNved tl-iem to be rer-zirst:e1v coupled to electromagnetic sorarces. I'lY:iS ~eatrare was used to eraable wireless control over btrt.}a the spatial guidance (using magÃaefic containers) as well as the delivery of naraolitr,r vo(uziir,s of chemical reagents. `I'i~e containers cati be gr:iidrwd in spatial p,a:ttems tE-iat: are aiot lii-raited by flow proffles.iÃa eotivetatiorial aiierofluidics, that is, downstream fto:m a channel iiiiet. Tt7e remote-controIled iiaric?liter containers enha:ncet~e capabilities of present-day xAcÃ-oflÃrid:ics by enabling spatially cÃ~iitrolted c(icziiica1 ~~eactioÃiS, miÃ:rofiabricakiian N.~iiÃbin capillaries, aald ola-dezYiatid localized de1iverd~ of chemicals to crxrlÃu.red ce(ls_ ~
100119_1 A combination of conventional mier~~fibriÃ:,atiran and self-assembly [1-'. G.
f,..eorrg: Z. Gu, T. Ko[i, D. H, Gracxas, J. Am, Ã;laem. 4oc. 2006, 128:
113"36-11 337; &
CJimi, T. LeoÃag, Z. Grxa, M. Yang, D_ AÃ-teÃiiov, I. N-1. L3tlr.riwa11a, D.
H. Gracias, Biomed. Microdevices 2005, 7, 341-34,57] were used tÃ) fabricate g0ld-coated, nickel naÃaoliier contairicr5 (FigLrrÃ; 1I a). To facilitate chemica1 delivÃ;ry, the c..orltairier-s were filled with a gel that was soaked in ti-re chemical reagent to be released (Fi.;tare 11 b).
'B.N~o gels were r_.rsed~ pluroraic.151 for general dry-release exprwr-iÃnentS
and po1y(Ne isopr~~py1acry1amide) (P1~IPAna) [T. Hirokawa, T. Tanaka, J. Cheni. Phys.
1984, 81, 63i9-6;>80; N7:. E lsl.am. A. INI6~~saved, Z. DogiÃ. J. Zbang. T. C.
Luberi41cy, A. G
YocJii. Phys, Rev. Lett. 2004, 9`21. 088303] for cheriaicai delivery in aqÃreoÃrs solutions aiid to living cells. P1r.aroÃaic is a water-soluble block copoIvraier bydrogel that scatteras at 52''C and is c:ompatiblc witl-i aiv.idc ran;Fe of chemicals [R
A1exa.ndridis, `I`. A.
Hatton, C:`oiloid SLart. ~i-ivsicoclieÃii. l;no;. Aspect. 1995, 96, 1-46].
Hydr-ogels based on PNIPAmIfi are thermoresprxrisive materials tliat are widely crsed in drLÃg delivery, because Ã1~ev ut-idcr~~o a structural tra.rasÃtion racar- the temperature rai-r,,~ of tl-re I-ILtmarti body [-H. Yu, D. W. Grainger, J. C'ontrolled. Release 1995, 34, ilittl?i; K S.
Soppimath, `i'. M. f"triiiriabliavi, A. M. Dave, S. G. Kurnbar, W. E. J-',LrdzÃraski, Drug Dev. Ir7d. Pba.rm. 2002, 28, 957-974]. This transition temperature, as well as the collapse kinetics ni":Ã'N1PAm, cati be altered by addirtg co-moticri-rers aild cliarigirig the degree of caoss-linkino.- [(~_ A. Stile.; W. R. Burghardt, K. E. Healy, Macromolecules 1999, 32, '7370-::379]_ Hence, PNIPArn i5 ati ideal candida:te for remÃaÃe-con trÃ?lled release to living cells at-id in liquid ri-iedÃ'a.
100 1.201 Once loaded, a container was placed iri tlic react.ion vessel of elioice arid could. be guided in ariv spatial tr,~jectory Ã.isi~ig a magnetic stylus. After guidaÃice to the desired 1oca.t:iori, a radio-firequericyr (RF) #Ãcld gcrierated by a 243 micÃ-ocoil wa.s directed towards t:lic coriiait-ie.r_ The pov,~er in the RIF field coupled iriduc[iaelN? Ã:o the metallzc container, thereby proclLrcing eddy cLÃrrcnts in the frame and heating it up by a Joule effect. It is possible to heat c;vÃ;i-a ~onma.gnetic metallic containers by indtictivc c.otiplirz.g, ari~.~l tlie heating meclia:riisrri is clitTererrt from tl`rat.
used to heat ~.~Ã~~lymeric magrietic microspberes. ;Sirice the coratairxers were inic.rnfibdÃ:,atcd, the electr'rcal characteristics cou(d be made reproÃiÃrc.ibIe, and the temperature coÃ71c1 be precisely controlled by changing t1ie irZciderit power. This rc;prodticibility shou(d be cÃ,`~l rtxa.sted with t1~e pozver needed for release trorrx polymeric magrretic microspheres, wliich cazi vary greatly ~ecatÃse of pol~'dispcrsiviN, in sizes and inhomogeneous distribution of magnetic particles within clrffererit macrosl?hc res.
1001211 Blv heating thc corita:incr, the gel encapsulated ztiitl`riri it sotteried (crr collapsed) and released the c.bemical at tlic targreted spatial location ~Figure 1 Ic.i. `l'he metallic cc>mainers are essential to obtain heating at the power and frequency settings tisc.d. No r:clc.ase was observed from tlic gel in control experiments (on expa,~sLirc to thc.
R.F radiation, btià :iÃà tlic absence of the container) bccatrsc of ric{,~Ii~ibIe dielectric heaiin_g at tE-ie kÃcqucricy arid power sirtttÃ.irgs tÃscd (see the Stal,po.it:ing Iriiormati(~n).
1Ã101221 The rettrote-coritrolled c:oa7talirei-s, rna:14_e it pi5ssib1e- to do cherrris-tr~, witli unprecedented spatial cotitÃ-ol .in 1-aardrtc3ri-eac.li reiorrs. 'I'o hÃg;hfigtrt this feaÃtare, Vve repaired a break oap in o:ire of two aqjaceÃit microwires embedded within a capillary;
the capilla), was accessible orily by hipt.lt aiid ~~t.ttpttt ports (Figure 12), The gap witliizi microwire 1wa.s repaired by remotely gtaidirig cntitaiziers to that site iti air Wi~;i.irc 1"
a,b3 and reirioÃely releas:ing first a chemical scÃisitizei= atid tltei-i an activator (t.tsitrg two wparate containers) locally at the site of the gap (Figure 12c). "1'he sensitizer and activator were titi ai-ad palladium catalysts, respectively, which facilitated the electroless deposition of copper. After scrisita.r:itrg atid activating the spatial rcgiwl withiii the gap ofmicro-wire lorÃIy (t i,gure 2d), tlrc eiiÃ:irc ca.p.illaiy was flushed Nvit}Ã a commercial soltÃtiott of copper su(fate (1~igtÃ:re 2e), <'11ttioi.ig}i both i-rucrow.ires and tlie wa1ls of the capillary wiere exposed to the copper su(1=aie solution, rtieÃaffic copper deposited oifly at the c}icmÃc.ally scns:itized atid activated gap in t-iricrowii=e I (t- igurc 12 f.). Electrical resistivity measurements c:crtifirmcd electrical continuity ol'microwire tac:ross, the gap. This result demonstrates the a,ttfllity of the c(irit~nor=.w for 1crc:ali,zcd clierzrical delivery and chemistry within capillaries arrd ~.~tEier small spaces. (iI
comparison to alrea.dv e5;.istinO; Ãiietliods of Ãnlcr~~affibrication in capilla:ries. J. C.
?~=1cDonald, ~ M. W(ritesidÃ;s. Acc. (::lrem, R.cs. 2002, 35, 491-=199;
MiaclotÃ, Ft.tnclanic.Ãita(s of Microfabrication, CRC, New Yorls. 1997: 13. J. A. Kenis, R.
lst~~~gflov, G. M. Whitesides: ;ScieÃicc 1999, ?85, 83a85]', this itxventioWs ziiethod is Ãiot limited bv . the geometry of tlie c:al,iIlaai= or laminar flow profileS.
1001231 ~secoÃic! demtÃnstÃ'atiotr. highlights the titility, o1't.tre rÃarÃo(iic.r containers in the r-cizioÃe-con t-rollecl, localized dirlivcry of sub-nanoliter volumes of chciiiicalw to specitic cells cttltuaed ori strl-istrates, Containers were loaded with I'NIPAÃn, soaked in a liv~.~:dead ('gaecnr`rcd) two-color fltÃcsresc.eticc viability stain [Irt-vÃt.t-ogen liv(-Aead stain product giiide to stalÃi cells 1ocally in a crjlttÃri dish aircl to t<t.~rify that Ãio Ãiet;i-i_stic cell death cscctÃrrc;cl dLariÃau cltet-nical release as a consequence of the heat.iÃÃws [S. Cc3r-On, S. Boexcb, C'. Maneschg, Ã.'. RacirÃ-layr, G.
.(~arfsc.~~, R
Klocker, ELar. Uro1. 20430, ;' ) 7, 499-5,04] or exposure to the R-E
Ã'a.diatit-11.t1_ 1001241 The L929 mouse fibroblast cells were cLÃ:(ture-d irt -35,-mtià well-plates wiih glass itilays and growrz to c.c~~ifltic:ÃÃcy. At the start of the rernotercreleas~ ex~erimcÃit, the ~prnwt:li media was removed and the cells were rinsed witli phosphate-buffered saJine to dilute tlie sertÃm esterase activity, tbere-by rriiriimizirig backg-round fluorescence. '~('o eÃiable reÃiiote release of the staiia, ati RF coil was placed below the plate directly tjÃider a coiitaitier, aÃid the coil was powered i.Ãp at '-2r3 W for tÃ-iiiÃltlt:e to collapse the. encapstÃlated PNIPAm aÃid refease the stain. Fliiorescent images were obtained 30-60 minutes after Ã-elease, toa.Iiowsufficient time for uptake of the stain. It is c.1eaa :i:rÃÃni the coiii"ocal fluorescence images (Figure 13) that the stairi was released loca11a<~.z~~~ ~~~ithiii a.Ã~.~.d.iuc c~l`-iess than 50~~ lt.i-Ã~. from the center c~#the c~~r~tal~~er. (t: cati ~
also be seeÃi that the cells exposed to the stain ha.d green fltÃcarescence, thtÃ;s indicating that they were alive, and Ãio red fluorescing or dead cells were obsened.
"('he results iÃidicat.e that neither the temperature used to collapse the encapsulated (''N:I:('':1m ri(ar the RF radiation c,a.used necrotic cell death. It should be noted that no lcakaye or spontaneous rÃ;1eas~. (that is, Ãio cell staining) was observed fÃ-uni the containers in experiments where Ã~~ RF field was a.pplied. Biocompatihility studies show Ã1o necrotic cell death occurs in the presence of the containers over 48 h.
1001251 Tn CcglIClLÃSitÃn, the metallic, self-assembled nanoIiic.r containers cwi. be titilized for rcniote>-coÃ}trol(ed rÃiic.rofabricatic~ti and cheniica1 cic:livery in hard to reach spaces. 'i'he containers will I~e useful in tabiicatiÃig complex and reconfigurable microarialvtacal, microflÃ.iidic, and micro-elecxromechartieai tvsterns. The localized rcnicitc: dc:1iven, of chemicals to cells establishes a methodology for r~.motely manipLÃlatiÃig thc chcmicai ajid biological micrv-eÃivirvnmerit for applications in cell engineering, tissue en4,~ineering; a:iid drug developÃiieÃit. Fina1Iv> the containers pro4.,ide aii attractive platform for the intogra.tioÃa of addit:ionai features of wireless devices (for emiÃnplc, frc.qÃ.eeÃic:v-sclective remote coÃitroI and remote coriÃmunication) with the delivery of tiancs(iitar volLÃmes of c.licmicals.
~XPER1MENT-AL 17E'll~,UL'-';
F'fkbr=iraÃic~~~ of the ilicroc0titainers:
1001.261 Brieflv, the fabrication process involved t.lre self asSemt7ly of a two dimeÃ-rsiorral (":D) template irrto the 3D cii.bic cnratainer. First, '?D
met.allic templates consisting of six square pc3r(yuS fiac.cs were pbÃ~to(ittlograpliica(ly patterned. A Secorrd layer of photolithography was used to pattern solder hinges ~ii the Ãxute:~-edges arid irI
beÃwcerr faces. `('lie '?D template spontaneously fotdeel iritÃa t.}re 3D
cubic contaiÃler wIreii it was }ieated 6rr a 17.Ãiiel) above the meltirig point of.'ttie solder liirlge5, wbereiri t(-ic surface tension of the ri-iolteri solder provided ttle 1`orce to drive Selircassemb1y.'f lrc final size a:trd porosity of t[ie 31) container was varied by patterning the 21) template alapro-pri.ately. In this experimerxt; containers were fabricated from nickel (Ni), a zrragrretic matcr-ial, to crrable rernotc tYuida:ncc. I'tie oiiter and ir-irrer surffices of the containers .:vcre coated witl-i gold (Au) to irac:rcasc E?iocoarpatib.ilit;:
ai-id decrease electrical resistance (low electrical reSistaiic.e, iracrcases t.lre skin depth for penetratiari of electrornagnÃ>tic: waves). The fabÃ-iUation process was highly parallel and large nLrml~~rs. of coritainers could be 1'a.bricated in a cost-effective manner.
PREPARATION OF TIJE C;ELS:
1'Ã301.271 PIr.rronic&: The gel ivas made by coa7rbirrirr : O.5 g of F68 Plrarcarrict (BASF) with 0.5 mI. of water. The mixture was sonicated for 10 mirrtF es to c2isurc complete mixing. Gelation c3c.cLirred after excess water evaporarecl.
1Ã101281 PNIPArii: Tl-ie Ã''Nl;Ã''Am gel was made I-roiir two stock solLitions, A and &
So1r.iiirYn A. consisted c3f.' I..6701. ~,s N-i.soprola?ilaca-ylarr-fide (1?Ni:l?Arrr), 0.0083 g N, 'ti'..
meth.vIerreb:Fsacr-ylaa:r}idÃ; (BIS), and 15 n-aI_, rarater. Solution B
consisted of 0.0129 ~ of amnionitini persult'ate (APS) and 15 mC. of water. Both solutions were vorte:~ed r.indl the solute d.i`solved. Gelation was <ac(i.leved t?t' mixing equal volurr-i~s of cacl~ soluficXi togetlrcr witlr 0.4 r~ (,v`v.) of Nv N, N`, N'yteta-aun~.~drvlenctbyl~.~rrediai-trirre (TMG[3) a.rrd oc.cr,trre(l w~ithin 5 niinErtÃ;s.
REMOTE dC-ti1DAN-(.`E;
1001291 Remote gtiiciaÃice was aclaaeved by inanipula:tir~~ a ~~~ag~ietic stylus below the reaction vesse(, ln ortter to redLÃ.ce t.tae f'ric.t.ioti between the comaÃnE~~~~ ~i-ici the su.r~ac.c of the vessÃ;1, the stylus was rotated alÃai1~,r the base of the vessel causing the Ã:Ãantainers to tumbl.e along the surface.
'i'HE 2D lf~U..N.11CROCt)IL SE'1'-UP:
[U0l.3()1 A ":D gniczoco.il was fabricated using phot:olithc~~~aptay on a f3r-irlted circuit boazd (PC~) as the I-',:F source. r1"be mici-ocoil was placed either below or above tlie contaiiiers at separation distances of approxiiitately 1-5 Ãnan. A c,lÃrrent at 800 MHz (RF) was passed t}irougli the coil to geiierate ar~ afteriiating zY~agÃletic., field iti a directiora perpendicular to the surflace of the c.oai; ata incident power ita ttae ran,)e of 1r7 Watts was usecl. The sufface of the coil was air cooled to remove any JoÃiIe heat gerteratec1Ãn the coil.
REMOTE R.EPAIR. OF A MICROWIRE WITHIN A CAPILLARY:
1.00131 1 M.icrow<i res (100 ~tm thick copper wi''.res, spaced 2 iiint apart) with break gaps of 50 ttm were fabricated ~~~i a glass slide using photolithography. Tlte capillary was formed usii}g po(yci.imethyI siloxane (RDTVt:S) walls that w~c;re scale:c~
against the glass slide (containing the wires) a:iad ariofliefl glass slide (the i=caot of ttae capi11ary ).
Seal'in4,~ was a.chie-ved bv plasma sa.Ãrface modi.f:ic:atioit of the :l'DMS}.
The capilia.t-y with the eanbc:cl.dc:d 3vire:s (f:=igÃarÃ; f?.A), wa.s approxiÃ-nazely l mm in widtla. and 1 .5 ci-n in length a.tid was ~.~taly accessible by the iiapLat a.nd ~.3titpuit ports it itti two eaids.
1001321 The sensitizer soaked E'la.Ãrotiic~gD gel was prepared by Ã:,ca.mbining 0.5 ztiL of sc;nait.izing soIÃiiic3a~ (Transene) witla 0.5 g of Plitronrc F68 (BASF). The activator soÃiked ) ~'1u.i Ãsni e ~ ~ gel was prepared by 7ng x,i.ta g 0.5 ml., of activating ;;c~l tati o~.ra (Transene) 0.5 g of pIar~.gnac F68. Prior to loading, eacii. mixttire was sonicated for 5 minÃites to enstire complete mixing. A 1 ~tL drop of eac.h of the mixtitre.s was pIaced on two separate Ã:ontai.riers ar.ad ttae solutions weÃ-c allowed to gel Ã.gvernight. (_14 hours). The containers were [bera cut out of the gel, to eiasca.r~.~ that the gel reniaiiaed only withiaa the cotit-ainer.
1 (1{}1331 After w*uid.ir.ag tI-ic coaztaiÃiers to the gap Mitbirl IÃUUr:arN~ire 1, WC ~t.iral applied 4 Wat-ts of power to tE-ie 2D coil until the gel softened and released the sensitizer solution from ilte container. T1Ãerà the power was t-edticed to _3.0 Watts -Lor l n-IiÃlLtt.e to allow sa.il='==:ti~cietit tizite for dift'us.ion Of tltC sOlÃ.atiOÃ-I through f1ic gel and to the surface ol the gap. 'I'he cl'aatine1 was tl'aeÃi flÃ.rs}iecl w:it}i water to reÃiiove the container aÃi~ excess geI. 'I`his process was then repeated, fk-.)r the secoÃid coritainer ti(led with the activaÃor solutzorl .
1001.341 .'4.ai electroless copper solution was Ã-iiade by rnixirig eomtiiercial solutions, PC clecÃreiless copper solution A arid PC eiectr~~less copper solirtion B
(both from ']'raÃiserÃe), irà equal voitÃriies. A .yz-int~e, wit.là a diameter of 0.9 mm was fitted with 0.9 znrri 11) ta:ibirig, 1'he (Ylkzer end of the tiibe was placed iri oÃ-ie openingof the channel. A
syzir7ge pLartip ((tAZF<.1.) was used to flow the platirig sc)Iution irito the channel aÃid over the 13rokeÃi microwires, A pulsatile flow was used to taciiitat.e the platirig reanon by maintaining a high local eoriceritratzori of copper ioÃis while <iilowirtg sufficient time t`nr deposition. During tE-ie e:x.perAiiaerat., the copper- plating solution was kept at.
45 C , REMOTE CONTROLLED DELIVERY TO LIV7.Nt. CELLS:
11101351 CoritaiÃiers werG loaded with PNIl'`'AÃti and allowed to sit overnigIlt, O1l the day of the cxpcr-ir.Ãient. the L.ive;/~~adO two-color fltrc3rescence stain (1:nvÃtrogen.) w:-Ãs prepared at concentrations of 0.5 cÃM Calcein AM aÃZd 1.0 ~iM EtlÃidiÃ,rrii hoÃnodiÃnor-l..
The PNIPAiÃi filled containers wei-c sa.ahr.a:ierg,,ed in tlzest.aiaÃ
st}lLrtion for 3.5 ilciLar-s prior to beginning the expei-iii:aerat to allow the PNIPAm r~t-n~le time C.k) rehydrate and absorb the liv(-~Idead stain.
10{ll361 :t.,9d~-~ mouse fibroblast cells (Sigma) w~,ere cultured and rrÃaiÃitairÃed tollowin;standaa=d c;ell. culture prc3toccais. The cells were cultured in 75 crn2 culture flask in 85% MiaiimEÃm Essential '~~Ã;ditrm Ek(gles containing L-<;~lutt.amÃne and sodir.rni bicarbonate with (0% laor;~~~ sera.trii mid s-Grpplementec1 witlà N1_EM non-Ã;sserltial ainirÃo acids and ;aÃ3ciitirÃi. pyÃ-taa=ate The cells Ã~iaiiÃtaiaied in ari incubator set to 37"C w.i-tii a. wa.ter-saturatec~ 5% C02 at--ai~.~sphere, 1_,929 cells were sul~sicultr7:red 2-3 times per week utilizing tr-yp;,in-1'11')TA arid seeding tEic Ãiew flask at a cieÃ-isity of =x 104 cells.?rn1;. 1'he seeding deÃ-isity was veritied by rerÃao-,>irig a sample of tl-re tiypsinized cells, stairring the cells Wittà t.Ã-y11aÃi b11ie aiÃd cÃsitia..- a bemacy-t.ometer to cociÃit tlÃe nr:irÃilaer of viable ce[ls.
1001371 :Kernote release experÃr-rietits were conducted in 35 rnrii well-plates witli glass inlays (I G1assabott.omsdislies inc.) for optinauni contoca1 microscopy.
Brietly, cells were seeded at a dezisiÃy of 250 tL of 2 x 105 cells:rÃi1:, directly orito the glass iÃ-flav <i.r7d allowed to recà .fioÃ~ 30 rYtin to prorYtoi:e adtlesiotÃ. 'i`wo rYii_: of g;Ã-owth ÃxÃedia was then added ajid the cells were irlctgbated for 48 hnurs to achieve a confluent monolaveÃ-.
1[){1138] After rerÃiote release, the cells were iÃ-iiaged usiny, a Carl Zeiss contbcat microscope. Br=ietl;;, the.Ãnicroscol~e was setup witb lasers and filt~rs, i~ecorni-fiended in the Live!Deacl,,x assay protocol. C;alceiri AM was excited at 488 Ãar~~ and ethidium horÃ-zodix~~er-i was excited at 543 nan. D-Ye up-ta:l5e was detected witli filter- eÃ-rbes of 1:31;' 505.5;3 )t] (for calce:iri in live cells) arÃd I.P 650 (for etlÃieliu-rÃ-r homodi.iiler iti dead cells).
CONTROL EXPERIMENTS
HEATING CHw'~RAf.:TERllS'li'ZCS<
1Ã10:1.391 To demonstrate control oa ei- heating of the nanoliter containers with tl-ic incident po3ver of tl-ifa magnetic field generator, a tc;mperatiive control experiment was condÃ.tct.ed, The sc:ttÃp of the experiment was similar to otir other RF
controlled release experirnents. TIw nrÃl_y di1l'er=erÃc~e was the p(aceÃ-nent of a Norirc:(teversible C)Nl:l:GA:I:,Al=;iEi.: Label (Ouiega `I 1:,-S series)urader- the nanoliter corÃ'[air7er so that it ccirel(l be heated witl-i good thermal contact. The i_ernlaeÃ`ai_ri.re of the CODÃrÃiTier sUÃ1~We cari be dedLÃc:ed color changes in the labels th,it occÃir at 38"C, 49'CY
60"C' arid 7VC
respectively. 'I'1-ic iiÃcideÃ-ià power c3t`Ãhe rnagrieÃ:ic field was iricr-eased u-rÃtil tlie specific label cliar34ed color (after Wai.t.ing .fo.r approximately .10 seconds).
1"'iw_ 14(a) shows the col.or- c:?ÃarÃ-~t:: in the Iabe1. oser` whicb the containeà w;Ãs lala~ed Fig.
14(b) is a plcÃt of the Ãc:.rnpertatÃarc measured b~:r t1~~.1~il~eis vs ihac iai~.~iehe~~t pÃ:g~n~c:r'. b~' c;l~at1;~irr~ t~hre ita.cident power, the heating could be precisely coaYtro_lhed. t;t slaoÃald be rroted t:liat the exact depends oia t-lre sh~eei#ie coa7taiiaer used and the experit-iieÃit (i.e. dry, or The color c(a.a.aau;e occurs on.[v Lirzcier the eorrtaiaier showiaY, tE-iat: the heafita4,~ is wet release, urrouÃadiÃag local. (B) A pIot of the temperature meastared usirag the color inclicat(yr s s label environment) vs the iracicleia:t RF power &Ilfri:I SI' A'1"lAi.: AND -NIATI:R-AI.T SEI..f:.C-1'lVITY OF REitlO'I'.I:s .It.I:si_EASI. FROM
TIiE CONTAINERS
1001401 To deÃrioÃrstrat.e tlic material arid spatial selectivities, the fo(lowiirg cc?ntrot expeririierrt was performed. 'I'wo eorrtaialers loaded wit:hr (''lurotridk:
4~;e1 (soaked ~.vith focsd coloring) were placed 3 ri-iraa a}iart: fro.in each other in a Petri disha. Aalotlier isolated piece c~fge1 (ia.ot eÃacapsÃ.ilated witbiax a ~~ontairrer) was also placed in the disla.
The Petri dis[i was aligned over a 2D xriierocoii at a distance of ? nirYr:
such Ãhrat in the plarae of tlie disla, the isolated piece of gel was aligned directly over the celater of tlle coil: one of the containers was aligned within the c:hrcumference of the coil bcÃt off'set from the cc:titeÃ~ by 300 ag-iel the second container was nri;;a(ignÃ;(i and lay outside the 21L) coil, ft'I-ie coil was hsc),wered up at 800 MHz. Only the gel wit.hila the container aligÃred avithrira the circumferetace of t(ae coil lieated LÃp aiid scafteraed at a power of 4.7 Watts. Even when the power was increased to 7 Watts, the isolatÃ;ci piece of gel placed in the region of highest field as well as the gel in the raiisa(ignÃ;d container remained unchanged. This coÃatrol experhrraeÃat demonstrates that the inclticti~~e beathÃ~g had }aigla spatial and material selectÃviN,. The ex~eriixaerat also shows that the metal used to tahsric;atc: the nanoliter cvirt-ainer is c ssetitial to enable the inductive he<atiÃag. It shoiald be noted that a Ãnar?ttetic matei'ial is g7ot Ã-eqtaired to fiacilit.at~
remote heating (the magraetic property is used morelv for spatial gÃiidance). Also demonstrated was the release from containers wihha iio Ni, i.e. c:oratiposÃ;d of copper / gold.
NO DIFFU'SIt)N OF CHEMICALS IN THE ABSENCE OF RF R.ND1AT.ION.
1001411 A control experiment was pt;.rtt}rmed to clem.onstr.,a[e abseric:e of dit-lb:;is.an of ihic I.1'':C /DEA:D' assay (i.e. dea7ron;t.ratC 110 tiPODUÃrrCOUs leakage of Ã;herTiic;als}
froiir loa(ie({ narrol.iter coratagne.rs, .ir.a the abscrrce of RF
.rad.iation. 'I"~ie e\~perinrerrt followed t}re test procedures wit1a: the exeeptaora that the RF was raoi iur-raec1 orta. "['Iri.s helped to etisure that the iii-tre frarrae f~'br exposure was the 5arrie.
Confocal rnicro;~~~py was used i:c) verif:y tEa.at: rro le.a.~~~ge, of tE-ie Live/Dead assay E-iad occurred frorxi the 1'~1:(=f'Am in the absence of RF radiation (remote heating) over the tirn~
scale of the exlaerÃraaerat (Fig. 1. 5).
Excrli../c, 5: Sur~ ~~c c> ?'ensiran Dravc-m Se1=FoNing Polvilcr~ra Fabrication ca f'patternect pofj~h~.~dr=a:
1001421 "1`lae t:inst step in the process irrvolvecl t:tac fabrication of 2D
templates c>orr-ilatased of patt.erriec{ faces and aoltier- lai:riges tlaiat woLr:(d eventually fiald iap inÃo 117 ho11o`v polyheclra. A polymeric sacrificial Iaver rr-iade of polyr~-ietlryl meÃIraca-ylate was spin-coai~ed oÃito a silicon (Si) substrate to iaci(atate a{ibseqtierat release of the 21) tezriplate4. A r-actaiiic seed laver was thea7 evaporated orito the sacrificial layer to create wafe;.r-sca:(e electrical contact ftar subsequent e(ectroÃlelaersitioi-i steps. The faces were p,atterrred using phr.rtcalitho~,s.raphy wid fabricated Uti11a1"
e::iect_rodepzgsitio.ta. Since conventional p}rotnlithograpE-iv was used to pattea=aY fiices, any arbitrary patierri could be .ine:orprarai:ed. Faces ccaFnprs:;ecl of eitlrer- copper (Cu) or rilcktl (M) were f_abricated;
c1:roice of n:retais was determined b~,,r cost, otcli selectivlty rn<ith respect to the sce-d lbiyer, ease of depositi t.a5 and the need fÃsr magnetic functionality. A 5eccrzrd ltaye.r of pIotolftkaosMrqplay v:vas tised tt) patiem the solder }ri~ige t. rlapl3tes.
After laingge patÃerrairag>
the exposed seed layer in the la.irzge re~lori kac3Ligaded by tEa.c faces was etcliec:i to cliscontlect tlac wrtieriy'.irrÃ~ seed layer oxrly between tl-ac faces, wfiile retainingr clectrical corrtit-wity Wit}r the rest of the seed laver at tlae face cc?rners. The solder hinges weru electr~~~eposit.ed, and then the 2IJ terr~jfla.tw was rc1ea`es:d I`roi-tr the substra:te by etcliirqg the reraiainirrg, seed lz7yfer-at3d ciissolvint, tlac sa.crifrcitrl Iaves. A tt,raapl:rte conrpt?sGt1: of six sqaarv faces, arrat-r~U~. in a c:raiei.ftrr-rra and held together by solder lairr~~~; ,va5 tisGt1: to form a crit)e Apart frorn the solder iaA between faces, there is no other tether. Self-folding was carried out .in a h.igh bc]ili_iÃ~,,, lac]iz3t so[vei-at, N rneÃlaylpyrsoi.idt>iae (i1;M1?), wlaicl3 tuas heated abave the iTicltirÃet poil-a. of the solder (-188"C) A small a_t-noua-1t of 44-5 RMA (rr)sin, rnfldlSf activated) f-ltix aA-us added to tlie solvi.nf to cleati and dissolve any oxide layers c~i-i t:hesoldvr and Ã.IiEreby erIsÃIre 4~c~c~~~ solder reflow.
13e:sigii t_onsiderati0ns:
1001431 lii tlic design, Ni was a1wavti used as the topmost surface layer of the face in contact wit}i tlie }iiiiges, Even for the Cta poIvliedra, the top ofthe faces were coated with a tltin laver of Ni prior to hinge depcAsitiÃ}aa, Solder does not wet Ni scÃrfa~~s w-ell, so the solder stays in the region where it is elc ctrodeposited. ancl does not spread across the eiitire surface of the face during ~~ldit ~~ (wliie[i occlirs wlien solder is in contact with Cti). When ttie Ni coa:tiiig was absent, we still observed folding, however the yields ,,vÃ:re pcscgr. The low yield was a resiift of the solder migrating away ~rcim the regions vvhe>re it was cl.cpositccf, thereby iiaaki~ig it very difficult to control. the vol~ime of solder in the }iinge region between faces (which ultimately determines tlle fizial ~~ldi~~g angle).
1001441 The design of the 2D template ir.Itimafely determines, the final shape and porosity of the polyhedra. S1iowti in Fig. IA is a typical -2D layoÃit of the tarcs ai1d hinges. AÃÃtodcsh AtttcsCAD 2005 was iÃ.SeÃ.l to generate the lavotit file L~~ed to fat?r:ie;aÃe two pltot.omtÃsk~ (one for the fzÃces, otte for the hinges). To fabricate a cÃÃbc, scfuar:e.fiÃc:e:~ separated b;~ ~gap, c, of 10--1.5% of the face d.in1ension, I.:, in Fig. IA, were t}'t.~ie;alk= U:~etl. SÃsrne tolerance in the gap width was observed, as the molten solder teiicls to draw t(ic fac e;~ laterally towards ea c;h other clLiring foldill.g. l:t shoitld also be noted that sir.aÃ:e the ~.~a}a wiÃIt:(i. .is I0--15% cxf :[.,, it was oftei-i tlle fniD.iITiÃ11Ti feature size of the pbotonnask and Iithcagi-apliv pioc~.`sk, e.g. i-~.~r .15 Lti-ii cubes, tl-ie requirred gap 3vidth of 1.5-2 ~um represented the st-nallest lithographically patterned feature.
1001451 :1n contrast with prior swrfzÃce tension-based seli=folÃ.ling, ovorlti, two types cal 1ii-Yges tVCT'e ÃISetl: internal ones betd~~eeii faces (folding hinges~ and external Ã.anes at the periphery of the t.'aÃ;es (locking biatges). Th.ae folding hinge witll}t (s:liÃ:AN'tt as W irt Fig.
11:3) was '?5% of L, artd the ht.ittw;e 1e2~~Ãh was 80r90`?i3 of I. If tE-ie folding htirlge lertgtlts were smaller the cÃrbes Ccst-a7ed were, iaot seal~d cotitplet.ely at the cÃarrters.
Longer htirtge lengths (>90%) were ut-iÃiecessary, sirtee rleigE-lborirlg hinges would overlap at t1ie corners. AdditiotiaIly, hinge letigt}ts of 100% were irto(artipatible wit}i the tabrica:tiora proÃ:.ess: tliese liirtge patterns Ã-esa.iit.ed in the Ã:,ot-rtp(ete removaI of the seed IaiJer at t:Ei~ peritiieter of tEic 2D templates during the etcb stop after Phtotnliih7o;.~rapE-ly of tl-ic laees. "1'his .retYtoy:t.l -forÃaYerl in art eleetrically discorttitl~torts ..,eed layer that prevented sul-iseqLient electrodeposition of t.hte hinges.
Re~o-w. of the folding ltirtges providecl the torque to rotate a~~acerrt faces. Lockxrtg hirtges that I-tad the same letigtli bLat half t_E-ie width of the to1ditig hinges played a secondary role in tlte foldino.~ (yl: the ~:I) tetr~~~latei thev as a stabilizing stop, increased fiartlt tolerance in folding, atid ertsa.Ãred a 1'atiat fold angle of 9V [ Sytits, R.
R. A. J.
~~,fic~rmd -evoorrtech. t. 1995, 4, 177-I841. Additionally, IÃsckirig hinges increased the t3=tcchtanictri strezt4xt~ht and se.ttli.d Ãhte edges of tf~.c t~st~f~>13~;~clr~a when ~-~~~o htali=<:ire~i l~~clÃ.ixt~;
11in~~es tÃtsed arid formed a. sin<~=te h~i~~e cotitait~ing the equivalent volume of a folciinQ
hinge. t=otdiÃtg was Ã:ott:tplete withzti-t seconds vdtest the 1oi<kistg lti''. ~,=es iitei kintl itFse~~ ~~~ttr each other. The fusiort oecut=ged as a resfalt of Ã1le tYtitiitYtizatiÃ.>tr of interfacial free energy beto~~eett the i-nolteÃt. locking solder bitt:;e on each face 3t-i~.-~ the surrounding lielrtid. 0-zt coolitig, the solder hinges solidified aiid the polyhedral st.ritc.tt.tre s~-as locked in to place.
Finite cleraietrt sintu tal.iotr t;
1001461 T:n orcior to better understand the self=assimbly, process, we perfo:r~~ed titiite.
e1enient sirt-tul.atiun5 crsig-tg the software prc3gr~n-i SLtrfac:e Evolver [Surface Evolver was developed by Kera Brakke -fi`oart tl-ie Susquehanna University Del:tartmertt cyf M,ithenra.tics. The latest Windows version v^.`26Ã;, Ãtpdat~.~ci.
Sehlteiitl:rtvr= .I T, 2005 was ttsr d]. Suafa~~ Evolver determines the tt-iirtimttm energy sutfacÃ; for a giveti. initial sLtr:face atid a set cyC physical ~~iistraints, suc4i as gravity, detisit.y, artcl sÃÃt~a0e tension.
The iterations for evolviazg a mininaÃa.na sÃga=f;tce are cctrttrctlled manually by -thie user.
Scripts were devÃ:lopeti to automate the task of varyin~.F parameters and evolving riireltiple surfaces. Sinau[Ãrtions pe.r.{=ornaed bave :irac:irzcled only f~~io acljjacent. SqrÃare taces, kield to~.~ether by a siriM,le solder fo(d:irawY hing;e, since this cal,tr.Ãr-ed itie essential frÃrietioti of ilie folelir~~.~ tÃinges, that play the ct-iiieal r=ole iri I~brmÃng i well-folded str-uCtUre. (hie face was asSLari-ied to be i^'axed, while the other was allowed to rotate frrvely around the solder hinge; tli:iS assumption parallels what was obServed in eNlaeriÃiieÃits. 'I"~o dete-r-mine ttie equilibrii-rm RoId a:ragle for a given geometry, we used the fiiol1owitig strategs- 1.~-i~.r:~t~, K arid Lee, Y. ~ =:
~'~=~~c~c~~ec~~rr~~s t~f Sf'.~1:; Sari Jose, tJSA.
1E398]: rrÃiniÃ~ia1 energy surfaces were g;eriera:ted t`or- arigles of rotatiori (out of the "D
plarie) between 0 (flat) and 120 (overfolded) in itiereriierital steps of 5 Tlle equi(:ibririrn angle coÃÃesponding to the g(obal mxnimrrm erxerg ~' was then tleterr~,iiretl frr~sn7 the rliiriiÃ-iitiÃ~i of the surface energy trend lii-ie versus angle plot, -for- a particular given face dr li3 E:11tiAoI:'l.
1001471 ShowÃi in Fig= 16 (B-F) are illustrations of ttie firiite eleriieiit simulaticyn f(yr the foldirig process. In ilie 2D teÃnp(a:te, the fo(dzrag hinge solder- iS iti the fc~rrYi of a "I'-sIiaped ri~.~E-It prism. On reflow, the solder IicILie#ÃeS arYt~ forrzis a Ã~ou-rided contour (Fi(I.
I60. DtÃc to the }iig}i interEacia1 teiisiora of the liquid solder (,..
481Ãn.1im) [White, D.
W. G. D-cxns. 1971, -~4~}t~7-3t~;~~~]. tliere is a st~'oi)g d~'ivi~~<~ fork:e to nunimize the e>~pased ifÃterl'aeial area bet-v~,teri tlle rYÃolteÃi solder and ttie srirrow3diÃ-r, t~ufdic licittid, 'i'his dririÃ~F.~ force cÃatayey the solder to ball rÃp w.hicb cesuÃlts in the rotation of atjjace at.
faces, 'T'he fold angle is primarily controlled by the solder volume. We observed evidence for this coeitrol M botl-a simtilatioais and experimental observations.
Different solder volumes gerieÃ=ated underfir?(ded (Fi~. 16 D. ( i ), correctly folded (Fig. 16 E. H), or overfolded (Fig. 16 F. 1) structures. A plot of the dependence of the fold ~iggle on solder -vol.ttme (generated by siÃiit.rlatiotZs, Fig. 17) shows that ttie fold angle decreases witls increasing solder vo1urage. ~xper-iÃ-nerltally, ttie solder vo1Ã.rÃiie Ãhat deterriliries the equiliba=irÃÃn fold angle w.as n~anipulated by coxitroIliÃig the lleight of tl-ie electrodeposited solder for a given lainge geometr),.
1001481 Sitice the scaling properties of the prot.e<sti were of interest, the gravitational potential eneqgy of botti the solder and t(ic faces were taken into account, in addition to t}ÃC iilterlac;ia1 IRÃ1-faCC Cr.aes-gtr ol'the soider. it has beeÃà shoo-~,-n by cÃthers and verifie;i-1 in our simulations tl-iat: Ãbe mkgnÃtude of g;rav:itat:ioaY,t.l e1=f=ects are esyeniiadly negligible compared to the iÃitcÃ-facial sLÃT=Cace eneÃaj~tÃtitii si-r.e-s become large (i.e. ttiÃià scale).
However, oa:Ãr inclusion of a o;ravitational ener(,w tet-tzi aIlowecl LÃs tc) determine the relative magniturles of each of the forces as the feature sizes were scaled up or dowÃt.
The fact that stÃrfac.e farces scale favorably witlt decreasiÃig size is aÃi attractive feature of stirtace teiisioÃi drzveii self-assembly aztd has the potential to provide widespread LÃtiliiy Ha tl3e assembly of microfabracated ia-iic,i-o aiad minz~sc:Ã1e siruct.tÃ:res.
(00149] t'Ãt order to determine the effect of size scaling ort the foldiÃlg process, sim.ulations for ':[) templates were performed witl-a taces sized ti`c?Ãxt the iÃ-iz~ scale to i}Ãc .ann scale for a fixed solder vc31ijÃiie. :(n cacb case, all diÃ-iieiisiotis (height, widtli, arid length) were l.ittearl~~ scaled 1~~~ tlÃc same c~~Ã~.st:~.tit i=act~~r. ri.it ~eriet=;~~~ landscape was ol3sen,ed (Fi4,~. 18) which drives the folding process art~l that tltere are different energies for differerzt fold atÃgles (for a ;;ivetà geÃaÃ~~~~rY. aÃ-id solder voluÃite). '1'he iÃiitial slope of the energy cLag-vcs isÃdicat:es the rnag;nÃtude of'tlÃc rotatiolial force of the faces and dctermzjics w17ether the f.'oIdiÃag process is spontaneous or not.. A
Ãtc(;at.iYe iÃlitiai slope (Fig. IS, 50 tim tc) '2 nini curves) restilts in a spz3ntaneous folclirig process while a positive iiiitial slope (Fig. I8, 4 itim to 6 Ãiim ctÃrves) indicates a non-spontaneous prcicess. The minima in the ctÃrves (Fig. 18, 50 tiiti to 4 mrti) around 100 are indicative of a stable, ecltÃilibÃ-iEim folded c.onflg),:uÃ'ation. The absence of a minimurn in the ctÃrvc for 6 mm, faces implies the absence of any stable folded configuration, i.e.
the two faces prefer to rema:in flat. 'I'}icse results c.ait be explained by noting that as the :~ize of the faces increases the weig}tt increases and gravitational forces begin to dominate ctÃmpaÃ-ccl to the surface tension forces in tlie mÃ-n size scale.
Hence, the initial slope of the ciicrg ,y landscape bccoiiies positive in t.hc mm rango. and the process becoÃiies noti-spontaÃteoa.is. At sÃxialler sizes, sa.irlace forces overcome gra:vitatianai forces and the process 1ieccgmes spontaneous all the way d~~~~~~~ to the Panosca1e. lt.
should be noted that in the siÃnu(ations, bulk properties for the materials and the solder were asstimed and c-AYUZ.ts such as phase segregation, inrcrn-Ãetallic formation, and.
d.iffiision `vi.tliin the solder ivere i~.inored, if't}iese atiaamption,,;.:
Et.olrl, it appears t}tat the self-folding process weatald work oÃ-a the i-iatioscale. For our stan~ard-eonletry, material Ã~ensities, atid solder surface [ensiOft, our SiRIiy~atatxa~~ ~~~ow tlie n1ax-IT11t1i-11 sponNne:,e-)Us t:c]Id.iaiÃ~ si~e to be L ;~= 1400 ~rrn_ S.iina.iia.taoais tilso slic?w that iii aii extreme case of zi Iow, surface energy hinge (10 Ã~yriesic.an; e.g. a la<lit.id polymer) aiirl tieavv faces ('0 g/c.an, e-.Q.
a dei-ise iaieÃ:a.l), folding is still spontaneous for poiyi-iedca as large 3s. 165 una. This implies that it Sliould be possible to fold structures witli taces cozripcasecl of alniosà ~iiy solid material ancl aYiÃla liii-ages ctsinposed. of vai-trially any liquefiable iYaat:Ã;'ria1 up to a size scale of ai'oL1t'2t'~ 165 pt17 for our particular ge<3J11et.[y.
Experimental results:
1001501 Experimentally, cubic poly[iedra rarigirtg in size from 15 pi7a up to 2 i7am were folded (Fig. 19). We }aave also been able to fi-Ad polyhedra of other sltapes. (Fig.
19C). Although we believe smaller polyhedra c:ait be fabricated, we(tave been limited by oter photolithographic capabilities. Below teiis of microiis, hinge gap width$
approach the sub-micron size :scale and alternative pattemitig techniques, such as electron beaiit lithograplay:are aequired to fabricate the 2D templates. Our theoretical simulations show that folclifl-ig of smaller poI~~~odra are spontaneous dLie to the large nlagtlittÃde of the surface forces at small size scales. Although silnulatioiis show tllat the fo(cling of polvhedra with large faces, i.e. 2 mm taces, is a non-spontaneous process, cxpcriiiae:atfi.kdlv w~. -k-vt;re zil_ple to -folt.l 2 ttam cubes. We rationalize tl-iis result based oii i-wo observations. Firstly, agit~itioii ti-rie to convectic~~i currents iri the lteat.c;d fluid occurs experir~~entally'. This agitation can provide the ini.tia1.
cltiviaa.g force tr.Y lift tae~.'s anaa-ginal.ly over the activation barrier for folding, Secondly, it shoL7ld be noted that `vh.ile ,ve proportionally scaled al1. size a=ari;rbles in the simulatiOD;; (C.g. a 2 MITi face was simulated wit~i a [bickness of 80 Pin}q it was tiot possible to do so e\perhnentally. Due to restrictions on tlie height of the photoresist and resolvable aspect ratios, we, electrodeposited a Ãbit.l:ra~~s of onlv 12 ~im for 2 Tt~iii ctibeti; the faces of the experimental templates thus had Ri substantially lower weight, i.nÃ:rea;:ing the threshold at which foldc~~~ ~eca:tne non-spontaneous to larger sizes.
AccotFrtti~g frai= this fixcd:f:ranYe tlYicl:~iess in otar si.nnuiation, .vc determined that the (str¾~e:,t size.to.r which the sei.f=.fioltli_11~ PFOcCss WOtIld ~vork ti~r tI-ac rnateri.t.is rt5ecl in our process is --:7 zmn_ A1tboug"1-I
we do taOt ea:t7eCt t-O ttse ',t lithrxgrapbic prrxc~ss, to fabricate ;trttcit.tres as larye 7 miii,, the process of seltrfoldin4x may still be refe~~~atit: ttà this ;~i.~~. scale, ~;~s~,~;~cirtJl~? in the i~ackrt~~it~~7 of elec:.t-rortic devices.
Tolerance of the prUtt'ss:
1001511 Wafcr scale patterning of the 2D te~~iplatc:s is highly parallel, O.g.
we t.~i.ck approximately 1000 (f.-l00 ltm) aaid I00,000 (.L='i 5 ~tm) w.D crttcÃftarrtxs (yt) a 3"
water. The fo1dÃng process is also highly parallel, and lat;;e nitmhers ot~:
2D templates can be foltlt;ci at unce. Experimentally, the t'olding process also appears to be considerably fault tolerant and we have otteti been able to aciiieve yields in excess of 90% a:tid t`alaricate large iittmbers ~~f polvhedra (Fig. 20). We lia~~e also observed that tc.aldin9 occurred even when hinge regibtty. was not pertectiy, centered across adjacent faces. Experimcntally, to increase fault tolerance, we ta.rgctecl ottt= solder voitime to result in a slighà overi'o1d (.._.100' of rotation trotn the hcarizortta(}.
Since we tt:sed.
locking ltiti<.~es, Ãl~is c~vert~~id etis~tred that the faces tiiet, allowing the locki~.~; hiÃ~,9es to ftrst.'. This increased the tolerance of the process [Svriis, RRA, J.
NIrc~roelc>c.rtrc~inec.=h.
1995, 4, 177-184] and sealed the cubes at the edges attd comers. Additionally, convection currents existed in the tiot scaltzticati clttriti{; the foldisig process.
J'hese c.otivection c<t.trresits wtt;witztt4'cf the 2D tc;ni~~latkas and. Ltic:rt;ases:i fo(ding, angle tolerance by encou.ra~.-ing the edges of tht:. fiact:.s tc) coll.idc., this allowed the locking soldei- liinges to fttst' and liolcf the faces to4~ether w=ith c,ottsiclerable strert4=th. CJacobs, H.O. et al., Science 2002. 296. 323-325-~
Gractas, I3.H. et a(., Sf-zr'nf'c' 2000, 289, 11 70=1 17?1.
C[1nCluSi {)Td S:
l001521 Irt conclusion, a surface tcrzsiota based fe~ldirtd; process has l~aectt presented that can be utilized to t`abticate utii:ct:E-ie.red, hollow patterned polytiedr-a wit:li a wide range of sizes from t:lie mm icp the nm. By leveraging wcl(-t;wtablÃsht;d lithograpbic methods in microelectronics, this faEaric.ation process provides ',i rOtrt.C
tO irI~:.Orptsrale precisely ~~igira.ecred monodisperse porosity, transistors, sensors, and otl-rer information processing devices ori the prxlylicdÃ-a to create "st-nart particles." ITsirig sira-11.ilati011s, we N~ave also demorasÃ.r=aicd iliat the Neling WOr.aid work e.vitla a wide range of t=ace mate:r.iak arid liquctial?[e, hin0es. also dernorastrat~s that the Lrti:liaatiora of inÃed'acial forces, which scale fa.vcaaably at small sizes, is a useful paradigra. for raiicro arid raaraatabri cat:i on.
E a.atn Ve 6: Spatialty Controlled Ch~.~tnist,~~~ ~ ~ing Renaotcty Galhl~.~c1 A'anof.zter GS'c=ale G~~ituhierv 100 1531 Along wit.la coraverational channel based rnic.rntllridic devices, sev'eral nanoliter scale claemical encapsulants have been developed, includin~. those based f~rl pc?lvriiers; gels, and litlr.rid drops [for example: (a) Lim, F.; Sun, A.. M.
Science. 1980, 210, 908a910. (b) Ch~ng, T. M. S. Nin Rev. Drarg Disc.ovcrv. 100", 4, '1l-235, (c) Langer, R. .3.cc. Clarrm, Rcs. 1993, 26, 537-42. (dyfice, J. D.; Sorig, H.;
Lyon, A. D.;
Ismagilov, R. F. Laai(ir~7uir 2003, 19, 9I27-9133.(e) Harnmer, D A.:
DiScla.er.D. E..
Ann. R.c,z=, I.Mati.r. Res, 2001, _3I, _187- 104. In cot rt:ra.st to the above organic systems, nzicromachinecl si1icori-based devices can have extreme prccisiori, high rcprodtrc:ibili.tv, excellent mechanical strength, good chemical stability, as well as the ability to incorporate sens.ira`s, sik;nal c.ond.itionfla;F, and actuating ttanct.iOD;; iD clOSC
prc.e:si.rni#-y oi- ori the saane substrate. H:txz~evcr, :;1:3 i-nic-rotnac1}iiacd nanoliter scale reservoir systems 3vith c:oritr-ollc (l porosity do not exist at the present time dLre to the inherent t.wta diriierasionality of t:(-rc photolithographic process tlaat is r,rsecl in conventional `ilicoia based r.a-lic..r-~rDachirling.
1001541 De;raaonSt.F-ated here. is the deveIopii-aent of 3D containers witla.
precisely engineered sLer-l'ac:c porosi~, and their utility in chemical encapsulation, gLricl~.~ci delivery, and spatially controlled c.hcmis[r~>. Brretly, the process irivr,>lved t1w photolithographic fabrication of.' a 2D metallic template with solder hia~ges (Fig. 22a).
The 2D template self-assembled into the 31) hollow polyhedron when it wa;
heated above the ri-ic;lti~~g point of tlic solder binges, whe.reiii t.EIC
stlrftÃ.c.C tc.DSiOTi zgf tllC tsl solder provided tl-ic force to d.r.ive Selt-asscn-ibly [(a) Syala, R.R:t.
Yea.[mall. E,M.;
Brig,lit., V.M , Whitesides., G.M. I 'NIEVIS 200-3., 12, 387 --- 417. (b) 1-lui, E. ;1; ; I-~ower R. =I ;Rocigers, M. R.; Uth :lnt. Conf MEMS, 2000õ 6021r607.{c} Gimi, B;
=1_eorig; rF; Gu, Z,;'r'ang, M.; Aa-teriiov, D.; 1;31~~ijwalia. Z.; Gracias, D_ H. Biomed.
Micr-odevice"005, 7, -341-3]. Containers 1~ave been tabrica:t.e~ I.N.1itla clit't'ereÃit shapes aiici volurnes ran~irzo t`rc~iia 230 picoliters to 8 tia~iat~likers (Fig . 22 a-d), I'he fabricaÃ:icat~
process was also 1-iigblv pai-a11el; containers of different sba~~~~ ~~ld Sires coulcl be fabricated in a single process run (i.e, from a single wafer, :1~'~~ 211 e-g).
When the process z~,as optimized yields ranged from fi0-90 o (yielcls varied fo~r difTerent sI-~aped caniaine.r~ depending on tE-i~ ~~~~~~~~er of fb(d:ing f=aces and ttie syrnmei.rN) for a 3-wa:ler, I":1le tii.~~or v.icld li.anite.rs itt the t~,br-cai:iort proc~~~
~~~ere t:lic phoioIit1iog-raphic fidelity in the reg:isÃav ot' hinge5 with respect t.o the tices, and t:l~e volu.me of solder if) t1ic hziiges ~Syms :R. R. A. J. MENrtS 1999, 8. 448-455. (15) De.ri.,. 'I'.:
Wliitesides. G.
M.; Rad}iakrishiiati, M,; Zabcyw. Ãi.; PreÃitiss. NI1'':.App1. Pllys_ Lett.
2001, 78, I7r5r 1777]. Since pIaotolltbcgCFraphic mic:rc}fabaYcatrc}n is highly precise, it was a.lso possible to pafterii one or more faces of the containers with monodisperse pores (Fig.
?22 h-k).
'1-1ie size. of tlie pores fomied was limited by the I.~1i~.~tomaskS Lised (wIlicli in this case had a resolution of 3 mici-otiS). Bv controlling the porosity it was possible to ent;Ãneer the rcaigeni release profiles as shown in Fig, 21.
001 '55,55~ fi'(ie c:onta.inerw were loaded ~isi~ig stereotactic Microilijecti~.~n with a solution cif a.:a;el (or po1~~r~~ea ) at~cl the chemical to be released. When the solvent evaporated, the gel r~inaÃnecl within tlie containers. The cheniieals were released by i.nuncrsing the loaded containers in a. sOlLItiOn that softened or cliswolved the gel (or ~.~~.~lvizicr). Siizce gds 4aiid polymers) are available wit:l~ a wide range of aolubÃIiq= aiid s~
c~lenfng temperatures, it was, possibte to manipulate the clieiiiica.l release rates tiSing different ~~~lventS an~ temperatures. The images shown i.n the paper were obtained tising containers, loaded with a block copolymer bydrogc.l (Pluronicnx-).
Release experinieiits were cl.oa-ita in a. water-alcohol 1.~asecl mc;di~IM (Details in the Sul]plernc.nitary SecÃ.ioazl..By varyiaig the relative poros.ity oia diffcrcntfaces of the container it was possible to get both isotropic. (Fig. 23:a.) as well as ataisotrol3ic. (Fi(T.
?3b) chernical release profiles. Since tlae f'abric.ation process was compatible with a variety of aterials, it was possible to l;al?r:icate nickel based containers that could be remotely guic1ed using Ãaaagtietic fields. A spatialfv controlled. (the letter G --any arbitra:ry trajectory is posslble) claciaa.icaJ reaction was demonstrated by directly releasing (~%.Titing) a p:H ÃzadÃcatoa- phezaolphÃlaalcin in a microwcll i~illcd witla an a1k<alinesolutinn (Fig, 23c). Direct writing was possible by iaaanipulating tlae plaezaolplathaleiat-pluronic loaded coiatairter usiaxga magnetic stylus that uasmt~~~ed tinder the micrc?well. It slaould be nÃfted: that while guided manipulation was clone usita.~.~ a laer~~iatietat niagnet, it is possible to use otlicr well-developed microcoil based raaag.aaei:lc ixaanipulation circ.uits, [Deng, T.; Whitesides, G. N1;
Racllaakriy tnait. M.;
Zabow, C~.; Prentiss, M.Appl_ Phys. Lett, 200 1, 78;. 1775- :I777]', to reproducibly control t:(~c moveraaeaat oi"Ãlae corztaiiiei-s and li~iic,e tEic spatial release o~t.'tlae chemicals witli arl}atrard. p;a.tte ta.s.
1.001561 Spatially localized chemical rcactioias were also demonstrated between mu.ltiplc nanoliter scale containers (Fig. 24 a-c). 4k"hen two c~.~titainers loaded Nvit1i copper sulfate and patassiLim hydroxide respect.ivelv were brou:;lat close to each otEier in an aqueoLis medium, a c1a.eataica1 reaction (to ~orixa copper laydroxÃde}
occurred only along the cciitra( line between t(ic two diffiasing rates; tlic reaction occurred nearer the containers witb the slower diffttsing chemical (Fig. 24d-f). These ex1,eriine;ntz farth.cr demonstrate that the spatial coaatrol over chemical reactions can be extended to more c.ornplek rexaction frotit~ involving mt.tltiple containers.
10015,71 -1n conclusion, as opposed to all organic cncapsia(ants, the comaÃners allow taizpreccclciated spatial coiatrol over tlac release of cheiiaical reagents by iTi rttic of their versatility in slaapes and sizes; aitisotropic l`a.ces; monodisperse porosity, and their ability to be gmdded in micrcgl'ltÃidic channels Lasliag magnetic fields.
Additionally, the nieta11ic containers interact witb remotc electromagnetic fields that allow thiaii. to bc easily d~.~ttactecl tza-id tracked (U5ing magneric resonance imaging, MRI).
ThLis, tlle containers proa=ide an attractive laIa1-Cora7a for e:ngi.tac.e:.rirrg rei-iiÃ.stc::ty guided, spatially controlled clae.mica.l reactions in rYiicrc3t lLiidÃ'c. systems.
Fabrication of tlac MicrOcOnta'sncrs:
1001581 .t1. 5.5 pznwthick Sac.riticia.i 1aver ot poly (meÃlaw 1.iaaettlacrw lza.te) (P'41:M,'L, MW: 996K) [Sigriia-Adlr-icli was ypuii oai a silic.on wa:fer.
On top of -lre PMMA-c.c~~ted waf:ei`, a 15.nna ad}resior3-prÃ.arnÃ:at.ir3g c}rr`crrTiiUM (Cr) l~IVer`
and a 100 rarii conductive seed copper (Cu) layer were evap~.~rci.ted. After tl-ae tlairl fill-a}
deposition, we spin coated a layer of Shipley 41'R.220 7.0 photoresist [;Rohiai and Haa.S, www.ro}uYahaas.ccam J, The tkrickriesS of the photoresist vva5 c.otatrtal:lec1 via the spin speed aÃid bv varyina, tlie number of c.oatiÃigs applied_ :1fter- a soft bake;
the resi,,;t lvas exposed to [iV light using an t:ltra pline Series Quintel ma.sk alip,.raer [QuiÃitel C't?rp., v,ww.quÃntelcorp.c.ona ] arad patterned using a ti`a.raslaarenc,v xnask. After developing the photnresisi; electroclepÃ3sitÃr~~i was used to ~.~row tlre rtaetallic t`ratnes of the znic.rocontainer-s with.ira the plaotc3resist mold to a heiglYt of 6w15 urta tdelsendirxgon t:lae characterÃstics reqa.Ãired by ~~arioÃ.~s applications). We used ccammercial electrolytic soIutions that contained the rr-aetal iÃ~iis of choice ['1"echaic, [ne., www:tec.bnie..cÃ-arar j tca e:lectrodelaoait dikTereÃat Ã-iietals. For the construction of non-mag aaet:ic coratairaer-s. Cu was electroplated, and for ma,xietic containers, :~:i was uSed_ A second round of photolitho.graphy was perforÃaaed in order to pattern the laiÃiges. A laver of SI'R'??U was sptira Ã.~ia top of the stibstrate aiad exposed tÃ.~ tlre hinge mask. WÃdeÃ-, irateà rral hinges were located betweet~ aqja.cent faces, whereas the t}ainrier, external liiriges resided at the otiter edgeS of the frames. Alignment muks were tised to enst7xe aligiiment of the hinges to tlac franie>s of the -2D pre:cLirSors. After tlae. Iringe patteraa;:
were developed in 451 Developer, the exposed C:Li (seed) a:Ãad Cr (adhesion) regions Ãra between tlle electrodelaosited frames were etc}ied using commercial etchants Ã:AP'~-100 for CLI and CRE-473 for Cr jTechnic. Inc., www.tec;hnic.comj). Tiai:'lead (60:'40, m.p. -Y
18~? T) solder [Tc:c:hnic:.q 1:11c,., www.tc:c;linic.c;om] was then electroplated iTitc:s the hinge regions.
The hei~.~lit of the hinges was apprc3xiÃiiately 16 ;~~i-ii. xkt`Ãer the solder electrocleposiÃ:iorl, the photoresist layers were stripped off with acetotae, the remaining Cu seed itid Cr adhesion laverc were elclwd atid the 2Drcp.reeursor teiTipiaÃe composed of rnetal frames connected with qolder hinu;~s was ai-nriiersed in NwMet~iv:1 l;'vrrolidone (N\11)} CSi~,~niar Adlrich, www.sigma-alcirich.coarij to dissolve the saci-ificial 1''NNIltIA
layer a:tid release the precursors frc~~ii the wa:fieÃ~, A1iproxiÃ~iaield 50 precursors izl NM:P
wer~ spread across a s~~iall crystallizaÃion disl-a aÃid a siYiaJl aaYioÃint of 45,' RM_A
flux [1Ã1diiim C'cirporaÃion, www.indium_com] was added to dissolve any solder oxides that may have.foa-med. The disli wa.s heated to 100 'C:` for 3 miiiut.es and then r~:~n-peci zip to 250 <,C' loÃ~ approximately 9Ã3 secoÃ-ids uÃit71 the solder becai-iYe ti-iolteÃi Durin4~: re{'lc?~v; i~- the sol~er .:~~et t~ie top 1aver of metal ofii the 2D precursor, the fzabnoation yields were Iloor.
Solder wet copper well but did tioà we Ni well, }ience for contaiÃxer,"Ii ith C'i.l frames it wa.s necessary to add a tliiii Ni laver to iriiprc~~~e yieids, WheÃi ;s6lder reffinved, [tie niolten solder at the Iii~iges asid generated the toÃ-qÃae to fold the 2l:3 precursors into 3.13 micrcgccgntainers. Upon cooling, the solder solidified anct permanently licld the container ~rwnea togethcr.
Ct~tititinei- Loading:
1001591 Two metbÃsds were tised tc? load reagents into the contaitiers, depending o-n the we;ttabili.ty of the c;hÃ;ia-iicaI reagent ori the c:orataines-. Wtieri the ctic:Ãi-iia;FÃ1. wet the c~iitairier weltq several boxc.s ivc:.re simultaneously loaded by im~~iersing t?ien-i in a drop ol='the chen:Ãie;al i-cagent. The solvent was removed by evaporatioii. This lc:ft behind the Pc?lviller [P( T011iocE~' F68, BASF, www.basif.t.oml soaked with the chemical reagent.
10016Ã31 The 5eÃ:or.ad nic:thod utilized t,,vt~ three-a~~s Newport micromanipulators [Models 460A & M462, uwu.neupor-t.c;otn ] to iiidepei}dei}t1y i:caritrol tlxc position oi' the microc:.untainc:r and the s'=rÃ~<~~~ [World Precision Insrr~r:~~~;r~ts, Inc:. :"~~~,~afi1T~~
Syr:irlgc;, www.wpnnt:.z.ora-ij. The syringe was ou[-fitted witli a 36-gauge ~-icedle [WPI;l 36 f_i-augc. Needle. wwwwp.ii.nc.com] t.o facilitate Io,Ãci.i~ig af the mic:.rOc:.ODtai~-lerS.
Cheniacal Release & ~eaclioaa Specatacs:
10016:11 lted I)ye (Fig. ~~a-b}: Containers tvere loaded witlr a mixture cnriipnsed of 1.6 rnl, i0.26I wYl F DX:{_` Red 40 [:~~~cCo-rri-iieA
& Co., lÃic., www.mccorxnick _corn] arrel an aqueous po(vtireric solÃrt-iorr Ã:.orriposed of 1.0 c, of P1a.rrozr.1c:F68 dissolved in 10 niL of water (18A ?~10) . A 2:1:2 (by volr.rrne) mixture of g1ycernl:ethanol:wate:r was used as ttie diffusion metl:itrrir arrd this medium was added to a small cliamber ccrÃrt.~~iiirig ttie loaded miÃ:.Ã-oÃ:.otxta.itier. The diffusion profiles were irr-rkged usizig a. stereozoom binocular rgzia;roscope.
1Utll.~"i21 caliyr-G uided 1'berro(pbt&ialei.ti-KOt=:l Reaction (Fig. 23c): 'f .lre indicator mixture for the pla~iiolplat(ialein-k.OH reaction was pr-epa:red by adding 0.25 mL of p}rertotp}rthaleirr solution {0.5 h; of ptr.errc?lpht.lraleitt [Frey Scientitie.
~~wv.v .:i=a~evscieÃrtitic .coÃ-ir] .i11 Ioorr-ii. ot ~~5%etlrarloi)toarr afli,ÃeoLispolyri-ieÃ-icsÃ~lÃtt:i(ari composed of :I .O g of PlÃrronic F`68 d:issc)lved in 10 ÃTiL -water= and loaded irrto a Ãrickel-based microcontairrea, The microc~~nÃairr~r was placed irrto a well of a tissue ctiItuÃ-e pIate, [;Falcozi(P)1~.1u:ltiwell`l M`l'issa:~e Culture lslate: 24 Well, arici a 1:1:1 (by vo[Ãr.me) g;lycer-ol- water: I ?~~~ KOtl(aq} mediurrl was iriirodÃtcecl irito the chanil?er. The micrcgccgrit.ainer was guided a-nd, controlled usMrw a 0.35 pcÃli lb., :1 1/8"
dia~icter A_(\iCo rouaid bar magiiet [McMastea -C"arr, ww-w-.mc:ma.ster.coÃr Ãj.
C x~~>~?ei ~ Il) St:al~`i'tt~> ~'c fr~<t~?t'~~t~ate fsl..)t~{ Rccic;ifon ff-ig 24a-ck L'uSC.)::~aq3 + 2 KC.)H(aq) K,S(.).,(aq) + C'Ãi(OH)>4s) 1Ã)Ftl.1.~"i31 The copper sulfate reactaÃit mixture was pr~epar-ed by dissolvirio 1.0 g of Plu.rorriÃ, F68 irrÃ:o 10 rril. crt' 0_5 "~=1==; Cu{ii}~~~~ aqueous yolÃrt:ior [Sit'rrra-Altl,riclr., z~ ~~,w.si_Qrr-ia-a1tir.ic.}i-com I arrd was loaded irrto a ri-iierÃsc:4sit.tairrer. The potassi111,11 hydroxide reactant tiaix[us-e was larelaai-ed by dissolving :1.0 g of P1uronit:. F68 fl-Itts 10 rrrl:, of 1.0 M KOH(~~) arrd was loaded into a second rnicroconta.irrer.
'f'1're micr-ocoÃ-ttairae~~ were placed in close proximity into a poly (dirxretlivl siloxane) [PDMS> Dow C:`orrr:iÃrg Sylgard(k# 184, www_dÃ~N.veorning.con) j trrier-owelt.
'f .lr.e znicro-~x-e:ll was fabricated by:mc~lding PDM:S against aii SU-8 p}rotoresist Ãn. aster: The ditfus'Mr and reaction Ãriedium was water.
1~=f3I1----Rcxcraicm (j_{g '4_ `.l'he iTtc{ic:a,tor ri-iixture for the p1terinlp}rthaleinwkOH i-cactioti was prepared by adding 0,25 mL of phenolphthalein solution to an ac1UeOLIs pÃ~IyrTtcrie sO(utioti eÃ7i-a7pÃ5s-cd of 1.0 g of Plurortic F68 dissolved in 10 txzl, water. The alkalirle ri-iixtu.re was prepared by acldirtg 0_3 mL ol 4M :Kt7H(aq) [Sigma-Alclrich. W-w,~x.sagtnaraldrÃch.comj to an aqueoLts polymeric soltÃtion composed of l,O g Pluronic F68 and 1.0 iTIL water. Two containers were loaded wit:ti the pli~iin11~lit:EialciÃi sc~lutioia azid one vvith Ãbe KOt:-:( solt-itiÃiti. 'The tkirec containers were tlw.ti placed into a 1'D:MS Ãiiici-owe(l, with water as itie diffusion atid reaction medium. 'I'}ie reactions were also imaged uSino a Stet-eozoom binocular micrc?scope.
1[){1164] W1ii1e the presetit: itivetitioti E-iaS bec~i described with re{eretice to the specitie evabod.itx~ents t}icreof it should be understood by those skilled in the art that various changes may be made atid eqttivaleiits may be substitÃtted without departing troti-z the true spirit arzd scope of the iriveritioÃi. :In additinri< many modificatiot~s may be zrtade to adopt a particular sittiat:ioti; ma.t:cr-ia(, compoyitiott of matter, process, process step or steps, to the objective slairit and scope of the present invention. Ali st.tc:li modifications are intended to be witliii t the scope of the c.laims, ap.1,e~~~ieci hcreto.
100011 Tl7B application claims the ber~efit of U.S. provisional application Nca.
60l81'.6,t)6-3r fil.ed 1tiiic, 2:33), 2006. Additionally, this application is a c:a,~ntintÃation-in-part of U.S. Serial No. 11,491,829, filed Tti1y 24, 2006, which claims priority to US.
provisional application No. 60/ 70(,903)õ filed JLi1y 12, 2005. `l".Iie etitirc co-ntcnts of these applica-ions ai-c incorporatezl,by i-cfei`eia.ce, hereirl.
C,Oi"Ett:NMF;N`!' RIGHTS
10tl02] This researc.la was stapported in part by tE-ie Nat:iotial Institutes of:1~-le'-flth (NIH P50 CA 10-3 ) 1. 75}. "I'lae gz ~'vernment of ttae Unitcd States mav have rights to this i~~~entic?ll.
FIELD OF THE INVENTION
10003] The prescrat invention .rc.lat.c.s Ãc) a iaiic.ro#~Ãbric~~ted nara.o-or Micro-sca1e particle for encapsulation and deliv~.~ry of i-naterials or substances itic:.tticl.ing, l:aLIt 1-10t limited to, bic31ogica1. incdia incluti.ir~g cells, pharinac.e::cabc:al agcnts, cornpositicarIs, dra.igz, ti;;sue, ge[s and po[y.mcrs contained vvitbiai the laci_rticle, wit1l subsequent release of the tkacrapeutic materials in situ, ri-iethods of aYaakitag the particle atad arletliocis of uszrag the part.icl.e in in. vivo or i~~ 'vitrca applications.
1;3At"11,,.GROUl~lD OF Tt-JE INVENTION
10004] Tn recent ~~~ms, advances in regenerative meclic:.-ine bave inspired therapies targeted at the ccllLilar lcvcl. TI-icse tlaerapies seek to implant cells or ceilLilar clListcrs>
manipulate cellular pat~~~~ays: aia.d target the delivery of drugs. For exwiaple, a wide range of cell liiae;s laztvc bec:ra csicloyed wit.liii1 semipermeable iiÃid biocornpatible irÃ-u-nobil:izaÃion devices that control the bidirectional diffijsiorà oi rÃ-iolectÃIes and cell release (R. P. LaÃira, J. L. Hayes, W. L. Chick, Nat.. :BioÃechnol. 14, 1107 (1996); t3.
Drive, P. M E4errÃatÃder, A. R Gascon, M. ~~ariÃ.aa, J. L. 1?edr-az, Trends irÃ
13iot.ec;.}Ãno1. 20, 382 (2i30'.); N. E. SÃnÃpson, S. C. Ciratit, S. J.
131~ckbalÃd. I.
Constantinidis, Bioniate:ria1s 24, 4941 (2-003.).). Concurrent advances in microtechnology baare revolutionized medicine; as Ã~ew implaÃ~table devices, ziÃicroarrays, biocapsÃ:ales aÃid mia;rcÃprcÃbes -a~~~ developed. "('tiese dev.ices have tacilit:a.ted ce[lular encapsulation, on-demand drLÃg release, and early diagnosis of dis~~~es, (:1_ T. Santini, M:_ J. Cima, R. Langer. Nature 397, 33-i (1999)s J.
Kost, R.
Lan4,~eÃ-, Adv, Drug DeliveÃy ~~ev. 6, 19 (~ 99t ); L .Leoni, '1'. A. Desai, Adv, Drug Delivery Rev. 56, 21 1 (2004}; B Ziaie, A. :[3aidi. M. Lei; Y. tFiI, R_ A.
Sieget, Adv.
Drug Delivery .(~ev. 56, I45 (2{304); A_ Desai, r! _ Wes"t,.M. Cotien, 't'.
:L3oiaÃsski, A.
Rampersaud, A.dar. Drug Deliveay :E' wv. -56, 166:( (2004); J. T. SaiitiÃii, A. C. R.ichards, R. Se}Ãeidt, .-N4. J. C'i~~~.-za, R. 1n.aÃigea~, Angew. C'beaYÃ. 39, 2396 (21000); L. .FireÃ~ian. E
Mahaji7as E. Broide, M. Shapiro, -[__. Fieli, A. 4terrlber; Y. Ke~lielmati. E.
Scapa, Gut 52. 390 ('U0)3~3_ In ~:Ã~ntr~.st to ~~cAi~raneric, l~~rdr~~gii, and ,cil-ge( based prw~cesses that have been tisÃ;ci for encapsulation and delivoryx coi Ãvet Ãtiona1 silicon (Si) based microfabrication has higb reproducibility, provides ÃiiechaÃiical and cliemical stability, a.iid allows the iiiccarpora.tiota of electrcÃiiie and optical modules witllizl the device, therf'.b~' facilitating wireless telemetry, remote activation and C:omTllÃ,iTl#cat#oll, in vivo.
Howevc:r. Si based microfal~iricdttion is inIÃerently a twti dimen<i~.~tia( (21)j process and it is extremely diffictilt to fabricate three-dimensional OD} systeÃ-ris using conventional microfabrication (M. Madou, Fundamentals of Nficrofabrication (CR.C, Boca Ratc}n, FL, 1997)), ~ 3D medical elevicÃ; has several advantages over its 2C3 c::ounier~.~saxt; (a) :-Ã
laqger~ emerna1 stirl`acc area to volumo ratio, thereby maxiiiiizilig ititeractions with tlie surrounding medium, and providiÃig space to moLrtit di.tTerent diagnostic or deliveÃ-y modules, (b) a finite volume a1k)~,ving encapsulation of cells and dnigs, and (c) a gc;ometÃ)~ that reduces t(ie: chances of the device being Ã;triclesiral.~1v lodged in the bt~~iy-1(1{}051 .I.n orae aspect of the larescrit iriveritjon, biocontainers (i.e., boxes, I-iollow particles) have becti fabricated by a strategy that conYbizies the advantages ot tfirce-dirr~ensiorraliiv with the desirable a5peÃ:.ts of Si-based microfabrication to facilitate the delivery of i:tier~apeLatic k(gents isi sitii. For ~xampie, the eoritairier-s are loaded rith ztricrÃ~~eads or cells e:rnbetlded in a gel, arid thus Ã:mi be used either in coqji.rnction with preserit dav immobilizatiori svstents Ã.ised, in cell eracapsÃ.l1atioii technolog, ~~, or tl~e~;~ c~z~
be used zndqendently. In a:riot.lier aspect, the biocontainers also caii be used fcar encapsulation o1':fiunctinnal cells witlairi the por-oras containers ior in vitro arid M vivo release of therapeutic agents with or wit(ioLit amma,rnosuppression. For exarnple, the cor-ltainers caÃa be used for ei-aeapsulatiori atid deiiveryot insulirx se-cretirxg cells for-iniplant-ation in patients wviÃh diabetes, for placing tLsmor innocula in ari:iriial riiodels where, e.onstra.iruna, cells ~,viÃltin a small re~fio.it is necessary, and ior-de.liver~, of tuaictioraal iietÃronal ('C; l2 cells. :I rt some embodiments, tlie faces of the container are patterned witli riii<:,roscale per-l:or-atiozts, allÃ3wint; control over per-#t-Lsiort and r-el~zise ol its contents with the sug-rortndirio; rYiecliLim. `I'be advarita~~;cÃ:uy attfibutes of the contaiiicrs are a parallel fabricat:ior~ process with vrwrsatilit~' in sizes and shapes; precise and monoel.i~pea=se strÃfirc:e porosity; and the ability for renwne guidance iisÃr~g magnetic ield.s. In another aspect, the coiitairzer-s of ttie present invention are easily detected a.rid non-invasively tracked i-ising CortvenÃional rt~agrletic resonance irnagin g (1LI~'~Iiand.
do not require the presence of a c;o-ntr-ast ~.~~ent, S'Li M_MARY OF THE INVENTION
1000G] The present iÃiveÃit:icÃ~i provides riaÃxoscaIe or microscale particles for encapsulation and delivery of materials or substances, xncludiÃxg, but fic?t.
l:izniÃed. to, cells, clrLigs, tissue, g;elc aÃ-id polymers cotitairied within the particle, with stibse~.~LÃeÃ-lt re:lea.qe of the therapeutic materials in situ, methods of #ab.ricatÃÃx~~; the particle by foldfiu-) a''D oreceÃ.rsor iÃito tkÃe ;3D parÃicle> arid tiae tÃse of the particle in in-vivo or in-vitro applica.tioris. In oÃ~~ ~~iibodi~iieÃit of the preserit iriventicÃtÃ; a three-di.metÃsiort-Ãat particle cÃ:Ãmlarises a mLiltitLide of Ã~~vowditnensioÃÃa1 faces itlat form a hollow, polyhedral shape a.iid containing a IillabIe center chanibe.r, whereÃÃl a.
size of tlle particle is niicrosc.ale or tiwiost.alc;. In wÃotlÃcr emliodimcnt the two-+:iim.ensiona1 faces of the particle are l.^~attemed wit.li 1.~eÃfibrations or pcares. .1n another embodiment, the perforations or pores are created photolitliograpliically . 1Ãx another embodiment, the perttarat.ions or pc?res have a. size from abotit tl. 'I nm to aboi~t 100 mic::rcÃns. 1:n another embcadim.entr the particle is fabricated troni at least one material selected I'rom. the group consistiÃig of a metal: a. polymer, a glass, a semiconductor, a11 insulator, and combinations t:}ie~~eof In another embodiii-aeÃat, the metal is copper or nickel. tn a.Ãiot.}ier embodiment, the particle is a Faraday t.a~,Te. In aiiother onihod:inient the particle is coated wit[i abiocompatible materia[. In arivlhgr embodiment, IEle biocompatible material is a metal, a polyÃxier, or a combiriat:icari thereot.
iÃ~ another iriil7odiriic:iit, the filla:ble: center chamber of the pa.rliclo is filled with at least one sÃ.ebstance comprising coiitents of the pa.rticle. In another einb~.~diinÃ;nt, perforations or poÃ-," in the iwo-dimeniioÃia1 faces of'the Pa.Ã-tsc.le allow re'lea.se of the contents of the particle. In another embc3dimentM at least one substance is ~a therapeutic ageÃit. In another embodiment, the the.rapeutic ageait is selected froÃn the {;~rULÃP
C0D;;iShÃ16Jr OI' a cell, a plÃannac.etaic.al. agent, a, c:ora-:tpos-ticÃn, a tiwstÃe, a. gel, arÃd a polyrnt,r. In another embodiment, t}ie particle is admlfl-iistered to a sutject and location of'ihe parfic.le HI the S1.1bjecà is non-invasively tracked by i31agi1ek rOSC1T1ai1t:-C i111a<q?ing.
I17. iÃ.I7.C3t.l7.Gr erÃabc.>dirnent, ttae particle is iniaged with negative coÃ-it.ra4t retiÃtive to back,F{rcawid or positive cLiÃ:ftl"cÃ.St relative to background.
100071 '~(`hÃ: present invention also provides a niethod of .fa,f3.rie~~Iti~~~
~1 il~~Oe-dinieÃ7sional particle comlarising a Ãiiiilt:it~ide of t~.vowdiaiensional faces that l`orÃ~~ a hollow polyhedral shape aiid containing a. fillable cetite-r= chamber, the method cornprÃsing tlie steps: (~~) fabricating a rnultatuele of Ãe.vo dimensionat faces; #.b1 patterning the fabricated ÃwoTMd'amerasioÃia1 faces; (e) patterning at least one hin4}e (ari the pattemed two ciimetisional face to tori-n a Iiiriged ed(;e; (d) J'oinitig a hiÃigecl ed~.~e of a first laatterried two dÃ'iiicrisiozial face to a hinged edgge of a second patterned t"i-o dimensional #ace, to torin a IiiÃia;ed JoiÃat; (e) repeating step (d) to form a two dimensiona.l precursor template having hinged joiiits between ad~acent two ~imensioÃ-tal taces; (t") lÃiluefiying, the ti'txiges of the two-dimensional temp1a.te using heat; <i.Ã7Ã1 {~} selt:r-asceiiif}ling the threercd:iÃvierisaonaI ~~t-ticie_ In another eÃrtbodi~~~~i-it, tfic hinges ol=_ ~tep (e) of the method comprise a material fhat can be liquefied. .(.ti a.iiot:lier ciiilaodimejit, the material is a. sold.er;. a. metallic allov, a polymer or a glass. In anot:h.er emlaodzmeiit, step (a) of the Ãiiei:liod 1~irÃber coÃTrprises the steps (:i) spiÃiriiiig a sacrificial fili-ii ori a substrate to 1 riYi a tirst Iaver, (ii) ~averÃng a conductive secoriÃl layer on the first layer; aaid (iii) patterning the layered st7bstrat~ by photolithography.
l:ii another embodiment, the particle has a size that is microscale or nanoscale. I~
another embodimeiiÃ, in ste~., (b) of the iiiethodt the ti.vti-dimensicrna1 faces are patterned witb pertoratioiis or pores. The perforations or pores are created photolithtagr;:tphica11~,. In another e;t7~~od:imentõ the perforations or pores have a size frtani about C).'1 iiin tca about 100 microns. In anotber einb~.~di~nent, the particle is a Faraday 100081 The presetit invention tt.irther provides a method of imagita.gy a tliree-dimcansi.ontil pat-tic:lc comprising a MtlltitLac9e of two-dimenwi~~ial fiaces that form a la~.~ilow polylicdra1 s[ial.~e aiicl coiitairiigzg a filla.blo center c~amber that Eias beon implanted into ~subjject, t~~e method, comprisiaxg, the steps nf` (i) loading the fi1:la61e center chamber of the paa~ticle with at least one substance to form a loaded particle; (ii) admiaiste:ring the loaded particle to the su~ae.ct; aiid (iii) noninvasively tracking the particle of step (ii) in the srIbAjec:t by magnetsc resonance imaging. In an.offier erribc>dinicia.Ã, perforations or pores :ir.a the bi-v'o-c.linieiiSional Eaces of the particle allow release (yl"tkae subcÃ.a.tac.e in the fillable c.criter ch.a.iTibcr. i~~
another etaiboditaaetai:. the at least on.e sub,,;iance of step (i) is ~thea~apeutic agerit. In atiotl~~~~
emliodymet5t,, the therapeutic a.gcnl is selected from the 4~rOa:ap corlsisting of a ce11, a lsharÃnaceLstical agent, a composition, a tMue, a <=,el, and a polya~er.
BRIEF DESCR[PTION OF THE FIGU RES
100091 Figure 1 is asclaeraaatic diagraiYi of the process flow used tot.a.bricate the :')D containers o1'tlae precetit invenÃz011.
100101 Figure 2: (A) Optical inaage showing a collection of containers. (B-DI) Optical and Scanning c(c;c:trosi microscopy (SE.NNI) images o:l'micrc~pattemed cuntailiers at ditTererct st~~~s of the fabrication proeess; (B) tlie 2L3 prectirsor witli electrodeposited 1`aces: (0 the precursor wiala faces and lainges, and (D) the folded container.
1Ã1011 1 -Figure 3: (A) ~ENNI iniage of a laolltiw; opGiiaf-tced coiitairier.
(B) SEM
image cif a cojataiia.er loaded u;itla glass ixaicroboads. (C) Optical image of a biocoÃataiiier loaded with MDAMB-~? j I. breast cancer cells embeclclecl than extra-c.cl:lular matrix ÃEC.N-11 gel. fl_7;y Fielea.se of the cells by immersioti of the container in war~i-i cell culture medium. 4E;1 Optical image of a container loaded with a ceI1-E~."M.-ag,-arose srispetision stained with the tltflorescent cell viability staiÃa>
CalceÃn-A:M. (F) Release of the viable cells from the container on itarnc.rsdon in warm cell culttire incr~ium.
10012.1 Figure 4: i4I_RI im~~~~~s of an ~peti faced (A;) noii-magnotÃc: CLi c.oiitainer a~id (B) ferromagnetic Ni coÃataiiicr. (C-D) 1~'itiitc e1~~~-nent simia(atÃon rewLilt; of the Mar rnagnet.ic field in the region of a L"Lgcontainer, iii the (C) xy aTid (D) yz ceia.t.ral planes. Ttic excitation cÃariaprised a linear polar.izecl 500 Ml-lz plane wave ofl V.'ryi, with tlic E. atid H fields in the z and y direction re;;pecrively. Tlie magnetic field cl.istoTtioias aaad the shielding effect caLaserl by t1-le wire frame are eviclent..
100131 Figure 5: M:I;, trta.ck.irag of a coniainer in a flLiiclic chaiiiie;t.
MR inlag ge;;; cal the container at tliftert.nt tinae points taken tÃ:iicler pressure drivc:n flow of'the fl.uitl.
1 (1{}l41 Figure 6: Optical a~id Sl=:~M i.7nzr_ges of the tl-iree steps rrsed to fabricate mic.ropattemed boxes. "i'he boxes sliowÃi iiavc approxiaratc dirTletrsioris of rxricr- ns. Frc~i-ti left to Ã=iglit- (a) The faces wer=e patterned using..-photolithography atid e:Iectrodepoaition; (b) Solder- hinges were ~.Ii~Y.t~ed rel~.tive to the faces usirx4;
phoÃolitho,graphy, etching and eIectrodepo~ifion; and (c) the 2D precursor was-lifted off Ã1~e wafer upori dissolutiari of a sacrificial (aver. When t.}ie 2D
precr.rà Sor was heated above the riie;,titig poirit of the solder, the siructr:rrc folded ilito a 3:13 cubic box [B.
CJimi et a1. voi. 7, p. 341r343, 200-5_I.
1001.51 Figure 7: Images of sorne defect modes observed: tt the solder }lei(Yht is not optiaiiiz~ed (A) uÃ-aderfolded or (B) overfolded boxes are obser-ved. (C) Incomplete etc.:hints of the seed laver usriallv results in faces that cannot fold (i841"),, because they are fu~ed together witii the seed Iaycr-100161 Figure 8: SEN1 arid optical images of boxes tilled wit~ (A & B) I?iuroziÃc hydrogel aiid (:') MUA-'-9:13-23 l breast cazieer cells embedded in e:xtracellular matrix (ECM) ge1. (D) =1'he cells could be rcleased from the box b;: pulsatile z~~~:itation in cell tt-redia.
100171 FirÃit=e 9: (A&B) Optical ii ~agos of 2D c(iÃl; fabricated tisir7g photolithography. By passing ct.rrrent thr-or,rgh tiie coils it is possible to generate a magrietic field. (C) The microbr~~ is placed alorig t..fae central axis of the coil in order to i.nduc:iivclv heat the bc3x, 100(8] FigÃire 10: Release of dye from a lo-idc;d. box r.apon heating.
1.00i91 Figtic=e 11: a) A scaaining electrori microscope irnage of an empty container. The containers were three-dimensional (31)) porous cLrbeS with a length of approximately 200 mm and a voILrmc of 8 nL. b) Ari optical microscope image of a coritai~ier loaded with a dye-soa-k-cd pl~roriic ge1, cf A scliematic diagram of t.lie ~Nperimental set-Ã.rp used to facilitate wireless micr~~ascaIe chemical engineering (Ãlot drawri to scale). Ccgr7tainers were manipuIfrted uSing a magnetic stylus (not Shwvn) and.
the contents of specific c~titaizie;rs were released by directiiig an RF
source tciwardS the container. In tbc; schc;matic representation, chemical Y is releia ;d from a specific conttzine.r; c;liciia:ical Ytl.aeri reac:ts wi~~~i chenaical N. in t.lic 4urro-und.ing r~-iediui-ii f~ca t`orni pt-nd uct I.
100201 Figtire 12: Optical imkges showing the reriiote controlled, spatially ]ocalized microkabgication witliin a cap:illary_ 'I'wo i-iiaci-owi.r~s (l and 21) were embedded within a macrnl='abric,atecl c.apallw-y (ca. l mm in diameter and 1.5 cm in length) aiid the capillar,v was aligned on. top of a ZD microc.oil, a, b) First, a container filled wit:li piriroiiic and soaked with the elicmÃcai sensitizer was guided iiito the cap:i11a.ry to t}ie site of the gap within wai-c 1usant~~~ a magnetic stylus, c) "l'~ie, cllemical wnsftizer was released bv re~inotefv heatiax, the sensitizer-soaked pluroÃiic gel that was encapsulated WithÃn the coiit.~~iier. '1'-(iis heating was achieved witb the 2D R:l?=' coil.
After sensitizing the gap, the first c.otii:aiÃier was removed, ~~econd colit.aii-icÃ- lvas guided to the same gap in microwire I. -i.tad t:fae actMator WIS released by heating the pluronic gel renioÃely, d) Atter activation, ttie second container was also removed. e}
The capillary was tlicii filuslicd with a commercial electrn1eSs copper-plating Solutiori;
chernica1 reduction (bubbles o1'thc: hydro4,en gas, a byproduct iti tt~e reaction, cari be seen) of copper sLilt atc to metallic copper, occtirred at the gap within microwire I. t) Coppe.r was deposi.ted otily in tl-iz gap $.~etwe~~i mic.rowire I, ~io c(il?per was depositc d.
in the ga~., f niiiierowire 2.
10021] Figure 13: Cell-viability asSe~smeiat by five/deacl fluorescent imaging of ca1c;eiai AM and ethidium homodimer-1, both released rei-note1~~ from the containers. a, b) Confocal iiua.gcs of the local release of the live/dead st-~n tci L929 inouse tibroblast cells. No red cells were observed, thus indicating tio nec.rkotic cell death during the aelease_ a) Transmitted light differential interference contrast (DIC) images sho-wing both the cells and the coiit-ainer. l;i) F(~~oresc:enr image showing on13=
localized cell sÃa.ining, 100*2 :1 Figtic=e 14: (A) Optical image of the color c.hang;e observed on a temperature indicator label placed under a nanoliter container, expos'ed to RF
raciiation. The color c l}agige occurs only under the container ~~owing that the heating is local. (.B).A plot oI'the temperature rneastared using t.lic coior-indicator Iabcl vs. the i ticicicnt RF power.
100:231 Figure 15: f_'csnfoca1 tiric:roseopy ir7rage of a tiatioliier c:orrtairicr loaded witNÃl?Nl:l?;1.i-zr gel soaked w.ith 1.1LEIDEAD<"~ assaw. 'i`1-le crpcrin-lent iotr?wed the test proceclÃÃres, v~=ith tlrc exception that the RF was not turned oti. --t'1Ãc absence of cell stainin4,~ surrounding the container (corÃxpare witb Figure 13) demcynstr-atcs no dzsccrr7a.lal.e c}Ãcrriica.1 release in the abscrrce of the RF radiation trigger.
fUtl24J Figure 16=. C 'orrrparison of finite simÃilaiio rand cxpcrimerri.al results for the self-assembly process, (A)'I~op view (drawrr to scale) wittr dimensions of the faces ~~id gap widt.}rs of ti-ae'?`D template used t.oselt-assemble t.lre cube. (B) Side v:ie-%.~~ of two ac.jaccrrt faces nf the crucÃt`orÃir (as tabricai:cd),with variables Ãased in tl-ic liriitc dement sirxiulatiorr. (C) Side view ol'at~iacent tacesat the oriyct o#-rci~~Nv r>f'tlie toldin }ainge. (L)-0 Firiite element snapshots sbowirrg (~~) urrcle~~folded.
(E) right-atigIe 1o1ded, aiid (F) overfolded fiaces. (t -I) Optical nricroscope iÃ-zrages of cxpcrirne.artall;; fabricated 200P.m ciibes exlribifing the Lsrlclcrfoided, right-angle folded, and overt olded faces. Note: Fig t.B-F are not drawn to scale in order to illustrate important (iiz-ncti5iosis.
100251 Fi~~tre 17: SimÃÃ1aÃiogi results of ti~e depciiclc~ice of the fold angle on solder vc-i1uriie The restilts deÃ~~~~iistrate that folding angle ca:ir be precisely eri;;irrecred by controlling the s(?Ider- vvIÃimc at the hinge.
100261 FigÃire 18; Normalized total cncqgy curves (finite c(cniciit simulations) plotted as a fiirictiorr of fold ang;fe for faces with Ieragt.hs r-arrgin:;
frorn 6mrn to 50 n ni.
The curves show that folding is spontaneous at sma.ll size scales with stable rafflirna.
As the scale increases, gravitational forces increase and folding is no longer sporrtanc~.~us (iiiitial slope cliariges t'rorrz riegative to positive) and there is rro minima preserit a:t 6 mm.
10027] Figure 19: (A) An optical image showing tiee standing polyhedra fabricated (c.xpc.rimeiita( rc.sÃilts) witl~ a. wide range of sizes all the ~~~av from '~~im to (B) 15 tÃni, and with ditTt.re;nt shapes e.g. (C) A ;;tlr.tare pyraniid.
100281 Figure 20: (A) Optical ii-iizt~,se of cubes With a .ratigÃ; Of StZCti I`zsrTtic-ld in lar~.=ertunilaers. {'1::3} lonri-ied in iniage of the otitlitied re~~iorl in (rl) feai.Ãtt=in~~ I00 pm cubes siitÃtic, oti top of and anion~..9 500 1mi ecgbes.
f00291Figure 21: Sync3pa.iS ol'toiding process.
100301 Figure 22: l~) Optical atid SE'~~~ images s17o-,.-ving the dÃtl'eretit steps (tlie photolitho4,~ral~l~.iÃ:..aJ1~' fabricated '?l) template, Ãe~;ist:r-~; of solder ItiÃ~tges and the folded -:D s[:rttcttire} in the fabrication of acttbic. container with one t7peÃi face. SEM images ot a(b) ct:tbic cotitainer ~vitb all opeti flaces, (e) pyrannidal fr-uStut-ti, (d) sqttarc pyramid with ati open face oii the bottom, (e-g) Optical image of multiple containerS
of dltTereait shapes demo:istra:tinia tl-ae paraltel tabrication strategy> {h=h.}
SEM itx~ages of cÃtbic cotitaitiers witl-i monodisperse pore sizes of (h, j) 5 atic.t=ons atid (;r; k) 3 m:icrÃ?tls.
10031] Figure 23: Optical images of chcralcal release from containers ( a) Spatially isotropic release of a dye from a container witb iclentiÃ:,a.l I?Ãarositv, oÃi all faces (la) AniSotropic release of a dve I'rozii a cÃ3ntaztaer- with aÃ-tisot-ropie porosiiy (five faces with ati array of 5 iiiic.roÃ-i porcs; ilic stxih face has a 160 i-tiicr-oti sized pore). (c) An e\arnplc of a remotely guided spatially controlled chetnical rÃ:;tction. The lettÃ:r G (for the Gracias Lab) was for-nie.t.i by the direct writirig of pheaiolphtlialein in an alkaline water---glycerÃ.~[ meclit.tm.
10032] Figure 24: Spatially cotitrolled cliemicai reac.tions. between multiple containers. (a-c) Reaction of copper sulfate and potas;ait,tm hydroxide in. an aqueous inÃ;Ãlit.tm resulting in the tortZ~ation of copper hydroxide along the central llrie between the Ã:..ontainers. (d-f) The reactlon of phetiolplatlialeiti (diffusing caÃtt of the two bottom containers) and potassium hydroxide (diffusing ottt of t.lie top cÃ}ta.ta:iner) in an aqt.tec=t.ts medi utii.
DETAILED DESCRIPTION OF TRE INVENTI+EON' 100331 The terms "hollow pat-trt:Ie." "box," ::contasner" and s.b1C}coI7tcltl7et`õ zEre used it1tf'.rcl1ai7~~eably f3ereii7 to i3lLili3 a three-dimensional C1bjjGct, i.c.., a rec:c;ptacle, with a hc.3i1mv inter'i.or- or an ititt:rlor capable of containirl_g sttbate~i-ic;e;;.
100341 I`lÃe term <;colloid'R or .:colioida(" as rase(i herein re::f-er-s to a substance tiÃrÃde Lap of asy5Ãem of partic[es dispersed in a c01Ãtia-ILiOr.aS r-freeliÃ:rm.
100351 Materials carà react quite difTeÃ-eÃitly iÃi the presence oI'aÃ-i extemal nia:grÃe't.Ãe tield. 'I'kiei.r reactic~ri is depe.rident orà a rart.inber of factors, including, but rioi: liÃYiited to, the ntaterial',, molecular structure, its, ator-rtie strLactÃ.rÃ-e, aÃid the Ãtet mao;Ãietic field associated witli the at.oms, Most materialS can be classified as fot`romagnetie, diamagnetic, or paraiii agri eti C.
1U03~"iJ The lerÃ-ii "diaÃnagnetic"' as used laer-ein refers to rYiate.rials having a very weak l'cirtti of magnetism exbibited only in t.lae presence of aÃx external magnetic field, which is tl-le result of chaÃtges irà the orbital motion ol electrons due to the exterrÃal rnagnetic t:ield. itidticed nia";ÃleÃ:ic r'laomerat. ÃÃi a diamagnetic mai:er-ial is very small and in a directioti opposite to that of t:lae applied lield_ 1-Nartipies of diama~.t1dic materials include, but are Ãiot limited, Ão, ct~laper, silver and gold.
1.0037:1 The terrn "1earornkgrteÃzc.,, refers to tnaÃeria.ls t iavirlg large and pca5itive Susceptibility to aii external riia(sÃietie field. Fer-romag-rietic rim:teÃ-izt.ls liave soÃxÃe ÃÃnpairec.l electrons so their atoms have a net ri-aagÃaitie moment. They e<bibit a Stront), attraction to t~~agnÃ;tic iÃ;lds and are able to retain their magnetic properties after the external field Iias beeri removed. Examples of firrr~.~magrietic rnateria.ls includel but are ntit 1'imited to, iroti: riickel and cz ~balt.
10038.1 The ter-rai "param~~~r-ictic: refers to materials lravi~ig &Ã small and positive sÃ.ÃSc:c;ptil.~ility to ni.a.gnetic fields, wliicIÃ are slightly att.racted by a rnagneiic fiel&
]'a:ratnawictic materials do not retaiii magnetic pr-opeÃties when the ext~.rÃtal tzeld ~is r~~noved. These paramagnetic properties are, clÃ.7e to the presence of sor~e unpaired electrons and the realignment of the electron orbits Carr.secl by t1ie extemÃ.-r~ magnetic field. ExaÃzit.~lcs of'paramagnet:ic rriaterialS inclrik, t~~Ã.Ãt are Ãi~.~t limited to, magnesium, molybdenum, and litlaia.rm, 10039] The terÃai "Faraday cag&' as t.r~ed hereirr refc.;.rS to a:ri enclosure designed to block the e#Tects of an ele.eiric field, wliile a11~),~ving free passage to magnetic fields.
(Sef E. M. PtÃreel1, Electricity and Magnetism, Berkele-y Physics CoÃ.ir ;
VolLÃ~ie 22 (N-l:cCfr-aw 1:=[ill, MA, 1985)B. SÃic}i an enclosure also is called a Fa.raday shield, Faraday Shie:[ding, Faraday ccrecsi; Faraday electrostatic sliie1dõ or shielded room.
100401 The tet=iii "gel" as used licreii-i refers to an apparently solid, jellvtike materia.l fnrnied :fi:rÃ~~~i a colloidal solution. By weight, 4e(s are niostly licli.-Eid. yet they behave like Snlirls, I"_[ie tenii "solution" refers to a homogeneous riiixture of one or more substances (tlie soleites) diss1 ~(ved in another substance (the Sc~lverit).
100411 '~('lic term "izidriÃ;tivc heating" as used bereiti refers to Ãtle process of heating a znetal object by electrognkgnetic iÃiduc.t-iori, where eddy currents ar-e gei~erated within the metal and resistance leads tc) Joule }ioatiiaz ~~t tbe metal. An induction heater (far ~~~ v proc.ess) coÃ-asists of an electrcima<wnet., throÃi<wh whiÃ:.1~ a hiwh-frecluency Alternating Current (AC) is passed. Heat mav also be (5enerated bw magnetic I-iyst:eresis losses.
10042] The teÃ'Ãn "magnetic field" as: L&scd Iicr~~ii r~~ers, tc) the re~ioti lri 5pac=~
surrounding a magnetic bodv or erÃtit:y; stÃc.l~ as a pertrianent magnet or a ccyiiduc-tar carrying a ctirreÃit:, w(iere ati appreciable magnetic force is preSCtiÃ.
Suc~~ a field is rcpresezit:cd bvinagriclic lisies of force. In an ciectromagnctÃc field, for exaiYipte, the magnetic field is perpendicular to the e(ectaacal field.
100431 The tÃ;rni "z~~~iielic field strength" or "magnetic field inten,ityõ
(<:ff') refers ÃÃ.~ tlie iiiteiisity of a ina.gnetic field at a given pÃ.~iiit. Mag-netÃc field streti:;t}i is, a vector qtÃatitity usually expressed in amperes per meter or in oersteds.
10044.1 The term "magnetic reso-nat7c:c imaging'. or "MRI.", refers to a noninvasive imaging technique that ~~ses the interaction bet~vee.n radio frequency pulses, a strong magnetic field, and an subject to construct images iti slices/planes from the nuclear magnetie resonance (NTMR) signal obtained fic}m the hydrogen atc}inS inside the suNect. The priiicip(Ã; behind all MRI is the resonance eqttatson#
100451 t1=:; B+, ( ~quation 1) 100461 which shows that the resonance frequency u of a spin is proportional to the n~~~~~~ietic field B~; it is experiencing, w~iere y is the gyromagnciic ratio.
100471 As used bc.rei.rr, the tÃ;rni <;mic:ro;Ã,a:lc" refers to par`tiÃ,lcs -tliat rrreaswret`rcarn about Ijinr or I x i C)'"; meters to about 999 l.irn in at least orrc dimerrsiorr. As used beÃ-c:iti the ter-iii .:.nanoscale" refers to particles tkiat measure trort-i i.l,oÃrt 1 rianorrietcr- 0r=
1x 1 F)`9 zrrcte.a s to about 999 rraÃroinctcrs.
100481 The term "ma gÃretic field ~radient" refers to a variation in the ma4}rletic field with respect to position, A one-dimensional mag-neiic field gradient is a variation with r-espect tci oric direÃ,tioÃi, while a two-dimcnsiozral ggradiezrt is a variation witli respect to two dircctions. `I'1-ic rr-iosi Lisct=ul type of g;radieÃrt in rnapetic resonance imatyirrg; is s a c~~ii~- dimensional liaxea:i- magnctic t~eld gÃ-ad.ient. A one-dimensional magnetic field gradierrt along the x axis 'Lrr a rxragrretic field, B,, indicates that the zrragrretic field is increasing in the x direction. I'lie symbols for a rnag;neÃic field gradient in t}ie x, v, aiid z directiorts are G:;, G and G.
100491 In physics, the term "riiagnctic rr~omerit" or .<dipole moment" refers to the pole str-ength. cit a arragrictic sour-ce mttltiplied by the clistarice betweerr the poles (p pd), arrd is a measure of the sÃ.reÃatYth of the niagnctic source. The rriao:rietic moment in a magnetic field is a mea.stire of the magnetic t1ux set LÃp by gyration of an clectroii chrirge in a mr~ggnz.tgc ticld.
100501 The terrii "micrÃ.~paÃtern" or .`r7ticropatterrreel" 'ati tÃsed lrerein refers to ar1y arbitr ar-v, two-dimensional pattern haviri<w microscale feat.tiÃ-es. The term nawpatternõ
or "nanopatterned" as uscd herein refers tc4 ~nv. arbitrary nsro-dimeÃisÃonal pattern l`Ãaving microscale teattzrc;s. Akcordirig to the present invcnti<Tn, the particles are paÃtemec1 with 1~erforatioÃrs c~~- pores r-aargirrg in size from abntit 0.1 niri to about 1.00 ml ct'ons.
1Ã1051 I The term "oscillatig~g magmetic field" or "oscillatory maynetsc field" r~~fers to a magrzetic field that pe-i-iod-ica11y increases arid decreases its irrtenwity, rlr, or wlricli othenvise varies over time.
10052] The particles of t~ic present invention may be in any polybÃ:drai shape. The term "polyhedral" as tisc;d lie.reiri refers to of or relating to or resembling a l,olyhc._dron.
The ierin "pulyhedron" refers tc) a three dimensional ol~ject boLindcÃ~ by plane po1ygoni or faces. T}.ae terna "polygon" refers to a mtiitisiticti <;cornet.ric figure that is bnui7ci bv matiy straight iiÃ-ics; such as a triangle, a sriuare, a pcrita~~~ri, a ~ie:~a~~~ri, a i~epta4ron., an octagon, atic{ the liike. FoÃ- cxatnpic, the particles oi't~ie-pr~s-etit i~~~~~~itiori mav be a cLibe or a tetra.liedrai.
10053] The te:rin "radio freqLa~iic.y" as used beaeiii refers to a fietitieiicy or interval of trequeiicies witiaiii the elect:roaxa~gnetic spect:runa Lised fibr com.munÃcatÃons, usually dcfiiicci as :~paiiriiiig fi-oiii aboui 3 k.Hz to about 300 t.i=HZ, w1licfi c~~tTesptlrtlds to wavclenubs of atjotit: 100 km to about I mÃii rcspectively, [0054] The term "radio trequeiicy tag" as tised herein includes radio frequency icÃeait'tfj cation (IR~~ID) tag:=. Radio-frequency identification (RFID) is aii azitomatic identification method, relying oii storing and rcmotely retrievi~ig data Easin;,~ devices called RF:(:D t~w=s, Ati :ft_[tID tag c.ati be attached to or incorporated into an object i=or the pur-pose of id~iiÃificatioii Lisiiig i-adit-~ waves. RFID ta~.s come in three geÃ~,erai 1,)wsrre, serrri -pwssive (also known as or crctive, Passive tags rec{u:irc iio iriter~ia( power soiirc.e, whereas s~i-iiiwpassive and active tags require a pt~~vea source, usually a small battery.
[0055] The tenii. "resistance" refers to a measure of the ~ie(zrec to wi`iicb an object opposes the passage of aii eicctric cLirrent as re~.~resclited by tile equation, RmmVs/I, where R is the resistance of the object (usuaIly measured in ca}iins, equivalent to J
s1~"}; V is the potential differe;nc~ across t.iic c3bjjec:tx usually measured in volts, and I is the curreiit passing thr.c~~igii the objjcc:t, usually measurc;d. in amperes).
1.005G1 The presence of a~iv sLibsta.iice in a inagnetic field alters that field to sc~~iie cxtent. The temi "susceptibility ettecx" rcfirrs to the degree to which a sLibstance't inherent magnetic moment prodtacc s polarization wheii placed in a ma:,rtctic tieid.
100571 ft'Iic tcrins "two-dimensional" or ::2I)'.' are Lised interchangeably herein to refer to a figure, ol~j ect or area that }ias i1eight and width, but no depth, atid is therefore t7at or planar.
10058_1 The terms "tbrcc-dimc.nsionaI;~ or "3D" are used interchangeably herein to refer to a tigtire, cgb_jtact or area that bas height, width, and ~lepth.
100591 I`lrÃ: particles of the prc5eait i.7averi-tiÃxn are fabricated using aÃ. least caric material selected :firÃ~ni tkie M,rotip consisting of a rTlctal 0-neaning aÃ-r clenicnt that is solid, lias a rxtet.aIlic lÃa-,terõ is malleable and dtrc.til~, aticl conducts both liei:t arid e1cctricity), a poI;;iiier, a glass (meaning a brit-tle transparent solid witfl iÃreplar atomic structure), a Seniicoridr.rctor (meaning ari e:leriieritõ ~swc1i as silicon, that is inÃerrnediate in eleÃ:.trical conductivity bet.weeri cor-rdLrctorS and insr.rlat(yrs, through whicli cozidrictiori takes place by mear~s of lioles atid electroris), and ari: irlsLilatcar (meaning a rxiaÃerial that is a poor conductor of 1~eat energy and electric.ity). They were designed as rnitaiat.Ãrre Faraday ca.<weS in order to facilitate detect:ian in :~::tR:l. The particles shield (r~~eariirig protect; screexi; block, absorb, avoid, or Ã?themise prevent the e1fec.tS of) tlie oscillating niagnet7c fields in MNM that arise fi"ori-r radio frequcricy (l~F) ~~a~lses aÃid t~~~.~wt~.et:ic ~iclcl ;.~r~.die.i~ts: in an itz~~.~~ir~.
; sequence. rl"~l~is sl~ieldirx4~;
occurs as a result of eddy ctgrretZts (meaning circulating currents induced izl a coiiductor aiioved tlia-nugh a ma>r~etic tÃelcl, or ~~l~icii is subjected to a war-virig ma#.~rietic field) generated in the -l:raÃiie cyf tE-ie partic.lc that induce a local magnetic field; which interferes dest.nic:t:ivcly witli the exxerrral mag.rret:ic 1=ie1d.
100601 In one aspect, the present invention describes the self-assembly of 3D
metallic particles frorn 2D ~.~hoÃo[ithogra:pli ically or eleetrtilithogral.~hica.lly micropatterned precLrrsors. The terms "ph~~to1ithography", .:pbotÃa-1ithography", ar "photolithographic process" refer to a. lit~ographic techiiique in which precise piatterns are created ori, substrates, sucb as mctaIs or rcsiris, through the use of photograpbically6 prnducecl masks. Typically, a SÃrbstrate is coated with a photoresist tri.m, wliich is dn'ed or hardened, and theti exposed through irradiation by light, strch as ultraviolet light, shiriiaig thruÃrgh tl-ic photomask.Thc urYprotecrecl areas then arÃ;
removed, tastrally through ctching, which leaves tlic desired pat:terriS. Electror~
beam lithography may also be used to create the per-f'arat:ions or pores.
10051] Tl7c. particles of tlic prcScrit invention are. Self-ti}lding and self-assembling.
The at least one hinge of these Stnactr:rres coriiprises a ma.terÃa.l, inclLr+:lir~g btit not limited to, a solder (meaning an a1lo~= formulated to have a specific melting point for u:+c J17 ]C1n1t7E_~', i'Ãletal;;), a 11#;tall9c alloy (mcs311Ã3g a Ã311)S.tui'e ['o31t'c713iÃ3.~w, twC? or 31C]'e nieiall:ic eieriÃcrÃts or gÃ-icta.llic aid nc3rlrneta.ii:ic clenicnts usually liÃsed together or di4so1v-in~,r irÃto each otkÃer .wlieti molten), apc~lymer or a. glass thai cai be tiqr.Ãetyed.
The surfiac:.e tension nl" the liquid hinge provides the force necessary to fold tlle "D
template iritn the 3 I) particles.
1.0062:1 In another aspect, after se(faassemti1y, the fiilable ceÃiter chamber of the particles of tiie present iiveitiori is available as a vessel for encapsulation caf t(-ic.rapeutic agents. As tised E-iereiii; the ÃerrYi 'the.ra.peutic agent"
refers, to any pharma~cuÃical agent, composition, gene, prot.caÃi, cell, molecule, or subst.ance that can be Lised to treat, control or preverxt a disease, xiiedica.l cc?xditioxi or disorder. The term "cornpnsitit:Ãn" refers to a rnia.Ãu.re of arÃgredients. The t:erri-i "
pharrriaceutic.at composition," as Lasc:d hereiri; reters to a c.oinposition, which has under gone federal aegulatory= review, "I'lie t.errii "treat" or "treating" iicltÃr;ies abrogating, substantially inhibitin~;, slowing or reversing the ~.~~c~t~essinrt of.' a coriditiori 5ubsÃantially arÃieI:ioratino clirÃic:al or syniptoa~is of a condition, and subyt:arÃtia.lly pr-eventirxgthe appearance of clinical or symptt~nis of a c:c}adition. The amount of a therapeutic agent that resÃ.rl.i in a thz.rapeutic or beneficial efTect following its admiriÃstration to a aub~jc ct, iÃicludirig humans, is a `'therapetirÃic amoÃ.ant" or "pharmaceutically effective arnouÃ1t".
The thera~etrÃic or beÃ~eficial etl'ect can be c.a.rriÃxg, rxiiriimizrrig, preventing ar ameliorating a disease or tlisordcr, or may have any. other iherapOrÃtiC or pharÃnac.c>trtical beneficial e6c;t,t. The terÃi "disease" tiÃ
">dis~.~Ãdr:*.Ã=," as r~sed herein, refers to ai impairment of healtli or a condition of abnormal fr.nct:icming.
"I'}ie term :.syrr.drorne," as ti:~~~i herein, ret'ers to a patternof symptoms indicative of some disease or c;ondit'itgn. The term "injur-v," as usc ki hereiii. refers to d~-nage or barrÃi to :-l sÃmeturc or tianction of the body calÃsed by an outside agerit or force, which may be physical or chemical. The temi "condition," as a.rsed herein, refers to avar-ieq= of health states and is meant to include disorders, diseases, or injuries ca.usCd by any underlying mechanism or tfisordcr, arict includes the prcir~iotiori of h"l thy tissues and cargaÃis.
100631 In so.me era-ibod.irnents, the fillabie center cl3arribcr of the particles can be i-ascd to encapsulate sLaclz therapeutic ag;ents as _pliarr-fraceratical agents or drugs, liairx4;
iis4ue. 4rels and polymers, tvliicli subsequently are rc:ieased in sitr.i. As used liereiri,, the t.er.an "pnlyrncr4J refers to aÃ-iatri.r-al. or- syr-iÃhciic compound consisting of long, repeated and somcti:mcs branched chains, built rip from small suburlits c.a.iled morlor7lcr.s.
Nattrr-al polymers incla.ide proteiris (polymer of a:riiiiio acids) &
ceI(Lilose (polymer of s~igar riiolecules}. I'here are mariy cxariiples of synthetic po(ymcrs, f00641In so.rnc embodimcrits, #i:znctional'==: cells +(c.~~ ;panc.rcatic is.let cells, ncurcanal :l't 122 cells) can be ericapsuiated t~~r in vib-o and in vivo release witli or without immunosuppa-ession. Sr.wh par-ticles cati be administered to a subject in xieed tl-rereofi by iiiic,roir~jcction, eithcr as a siÃ-igle biocoritairicr or as :1. ;rou~~
of"biocontairlers aÃid are useful for imaging, diagnostics-, and t:hera.peutics.
100651 For exaniple, in oae embodimerit, f1ie iritericars of a multitude ~f particles were filled with cells that were cziibedded in agcl. I'hese cells could be released by imrrie.a=sing the biocontainer in ari ap}irc3pfiatc so(verat. `I'he rYiagrictic resoi-ratic,c (MR) images of the pailiclcs embedded in tlr.lidic media srr~gest RF shielding and a sr.e4ceptibility effect, prcg-v-iding tharac.tera ;tic 1-aypointc:nsitv (+:larknc ss) wiihin tbc particle, thereby alk-nving the particles to be easi ly detccted, 'I'lii-, dcmoÃistraticrl is the first st:ep toward the design ~~f "ID, micr~~laatterncd, rioriairiva.sively trackable, encapsulation and delivery deyricca, 100661 fi'he present invcntiozi provides a three-dimensional particle i:omprising a plurality ot ovo-dimensional 1`a.ces capable of self-folding to form a holimv Ãnteriar, wherein asi~e of the particle is mic.rosc.a(e or nanc}sc.ale. The particle preferably ranges in size from 1 nna to 2 mm.
100671 The part:iclc flrrtltcr comprises at lea-st oric hinge, whÃcli may be ccrriipriscd of any liquifiable mat.erial. For example, the hirage mav be apolvriier, a gel, glass ar a m etal .
1Ã1068_1 The parÃicle of the present inverit:iori Iias any shape, but prefirablv has ster:fac.es fc?r'ming apolyhtadra( shape, sr.rch as a crrbc. The particle's two-dimensional iaces are pat_temed with perf1wations oi` pores. These:: perforations or pores may be created phntolÃt:howrapl-iÃcally, electroiithog;raphÃcally or using electron beam lithography. These perforations or pores 11az~e a size ranging frorn 1bOÃJt 0.1 tirl-l tÃa about 1crn. :('ret:erably, these perforationS or pores have a size frori-i about 1.0 rti.m to abaui :l cm.
1.0069:1 '1'he liai-ticte (container) of the preseiit invention may be fabricated from aiiir trtaÃerial, laut preferably at least c~iie material selected 1=roiti the group consisting caf a i-zietal, a polymer, a glass, a SemicondÃtci:or, an ii-isÃalator, and co.mbinations ttiereof.
'I'lie paa-ticle n~ay also comprise active electronic or sernicondiÃctor ecyiiipcyrietits suc1i as transistors, sensors: aetuator:=, lig}ii emitting, diodes, photodiÃ?des and solar cells. If the particle is metal, such metal iiiay [~e copper oà rric_h..el. In otie embodiment, t~~e particle is a. Faraday cage. In another einbodi.itient; the parti~~e a~av !~e coated with a bioeompatible material, such as a metal, a pol~iner, or a Ã:,onibination thereof The panic1~ ~iia~~ further be associated wil(i abiowrtsor.
1[)070] '~(`he particle may#iirtl-acr comprise at least oÃ~~ substance, sue~l as a therapeutic agent, eiicapsulated within the paÃticis;.. The therapeutic agent nlay be a cell, zt chemical or biological a~~~.ait, a pharmaceutical a.gent, a c:oml?ositi(iri, a tisstÃ.e, a gel, aiid a polymer. In certaiii embodimerits, perforations or pores in the two-dimensional faces of the particle allow release of the contents of-the particle.
10071.1 The particle of the present invention may, be adminis"tered to a subject. 111 such ~~~i etnbodiiiient, the loca.tio~i of the pw-ticle in the subject may be non.ainvasively tracked bv magnetic resoÃiaÃice imag-ing or CAI, scar~ (Crl"). 'l`~e particle may be imaged with iieS.sat:ive contrast relative to back<wr~~~~nd or positive contrast relative to ba[:kgÃ"Z713t7d.
100721 liz aiiotlier ~rnbcdiment of the invention, the particle additioÃially con1~.~rises a radio frequency tag, wherein ttie sa.ibstata~~e may be released upoÃ~ the particle's expoSÃirÃ: to a pro-Selected frequency.
100731 In a t:r.rr't~~~r embodi.F~ient of the partic:le Ã.xf the rnvel-1t.i011, the substance rr~ay be released Lipnri the particle's exposure to electrori-ia(Si-ietic radiation, wI-rich r~iav be tri~.rger ed r-etrioiely. The eleetrcsr~~agcic:iic radiation i-a7av r=arigc from lK I-iz to 1Peta Uiz 1U074J In a:fiÃrrÃber cgnboclirnent of the particle of the .invent-iorl, the srrbstarice may be released tilaort the particle's exposure to indLactive 1~eat:ir~~~;_ Sueli iriductive heating niav be taig*..~ereci reriiotely.
100751 '~('l~e preserzt irivcritioÃi also provides a incttlod. o-t.' fabricating a tklree_ dinier7sional partic[c cÃ3rzilarising a Ãiiiilt:itLide of Ã~.vowdiinensional faces that )`or-r-fi a hollow l,olyhedaal sliape a:rid contairiirig a tillable center chantber. This txiethod cor-aprises, the steps, (a) fabricating a multitude of two dimensional faces;
(Y) patterning the fabricated t~~~Ã~-dir~ieri~:ir~i~.al #ac.es; {~.) paÃt:er-riiri~.~~ at least Ã~rie I-rirt~~e ~ari tfic patterned two dimensional fiace to form a hiÃi4>ed edge> (d) joinint; a hir~~,~ed edge of a first pattemrrd two dimensioÃia.1 face to a birx(;ed edge of a secand patterried two diriierisioiia( t:ace to torrii a kiirigedjoizit, (e) r~~~~eatirig step (d) to fori~i a tNvo dimensior7al precursor teÃiiplate 1~avirig hing;ed joii-iÃs bctweeri adjjacent t,~~Fo dimensional faces; and (f) lic:ir.rel=='ying the hinges of the t.~vc>-dimertsion;rl tr~niplate usint;
heat to iiiiiizite selrr#'-fulding. This method a.(1~)~,vs the pal tic.leio self-assemble.
100761 In one, embodiment of this niethod.t the hinges of step (e) comprise a mateCiaJ that can be liqa.Ãetied. The mat.ctial rnay be a solder, a inetallic a.llo`, a polyn:rer or a glass.
10077] In mothcr e.m$.~odimerit of this motl=aod, step (a.) ftirther comprises tl rc. ste.ps:
(i ) spimiing a sacrificial film ori a substrate to form a first (aver; (Ã.i) layering a conductive second layer cgri the first iayen, and (iii) patterning the layered substrate by phoCo1 i ihtagr;:tphy.
10078] liz tlicse met1iods, thc particle has a size that is riiicroscale or rraÃlowcaIe and may have two-dimensional faces patt.cnacd Nv it.la perf~~arations or pores, whictt may be created p}totolithcggraphically, and may vary in size from about 0. 1 nrn. to about 100 niic.roiis. The particles of tliese r~ietliods may be a Faraday cagÃ;.
100791 '~1'1Ãe invention further cumpz.ises a m~tht)d of irÃ-iagiÃig a.
particle of the iÃiveÃa:iorà that }ias beet-i igi-iplaÃited it-iÃeà a Sr:abiect coÃ~ipi-ising;
the steps of: (i) Irladirig the hollow irÃterior of the particle -witlà at least one substance tE3 form a loaded part7cle; (it) adrzÃ.ir7:ist.ering the loaded par-ticle to the su~ject; and (iii) rionrcinvasively tracking the particle of Ste}i (ii) in the SubjJect $?v magnetic t'esoriariee :irnag-ing.
1n one ernbodirlient, the particle has pertoratiotiS or pores in its two-dimeiisioria.l faces that allow release of the sulasÃance W tfie hollow iriterÃor. In orie eÃiibodiÃiierit: at least oÃ-re substance of st~-p (i ):is a therapeutic agerii. 1'he therapeutic ;Ã.oerat may be a cell, a pharmaceutical agerÃt., a coÃtiposiÃion, a tiqsue; age[, arid a p~~fymer.
~00S0:1 The meti-aocls of the invention also comprise a met}Ãod of treating a condition co181p1"ÃSltl%M introducing 9.ilto aii a.t1.tI71al til I1eed of tl"ea:l'rtlellt at least. C)ile particle of the .iriveritioti encapsulaÃ:iracy :Ã. 001Ã1posit:io11, wlier-eitr t1le composition iS
released through orie or more pores witlain the par-t:icie into the inammal in an amount sufficient tc) treat tlhe coridition. 'I'l~e pharmaceutical coÃ-zipÃaSitÃoÃi rii~~~ be contained within one or Ã-iiore m:icg-e~~~eads. In one embodiment ot'thi~ inethod, the coridition is diabetes, ar7d the composition is one or more irlsr.llin-Sec:rctirr~ cells.
1Ã}041.] The invenfioÃ-i further provides a method for imq ~ing a ~,~tii;1 of f the iÃiveÃiÃion that has been introduced into a ma.nimal cor-ri~.~rises using ma~,~netic resonance ima-ging.
10082.1 The iriver-itivr~ further provides a method for= rargeting the particle of claim 1 to a. cell within a sLÃ~jec t eomprisirig the steps of: a) a.ttaching t(i the particle an antibody aga:inst aii atitigeÃ~ specific to t(ie cell; and b) introducing the particle into the mammal, whereizi the particle is targeted to the cell, 10083_1 1n aÃiother aspect, cells withiii or proximal to implanted putsc:le;
of the present invention can be irnaged by MRI to evaltaate tlÃC efficacy of the implant and the condition of the encapstÃlated cells, 10084] The invention also provides a ii-acthocl of dc.Iiver-ing orte or more particles of the invention to a suL~je:et, wIie;reirà the particle is pro<Trami ~cd to remotely rel e.-Ãse, one or more reagents at a.n~= specitit, time and at any specific spatial location.
(n. one Ã:rribodinienà of this niethot{, the particle is i`eiiaotely guided a~id Una{,~ed tisir~g 1A.RI car C"1`.
100851 Also pr~oviclecl is a method czf'reieasirtg a Ã:.ontrast~~,',ent from the pi.rticlc: of t(-ic iriventioti or of pg-Ã~vidisig contrast to allow ?,~I:IU or C,.l' iniat;int; of its coÃiterits or of stabstattces`Niit_[iin its vicitiiÃy_ l.00S6:1 A metliod is also provided for conductir~Ig non-invasive biopsy c?f znic;rosurgery, coiiiprisiÃig dÃrectin~; the particles to a site within a si:il~jec-Ã using reÃiiÃitc r~~eaÃ7S, allowing the particle to captiire i~i-ic or ÃTloÃ-c stabstances i=re~i-ii itle site, aild obtaining ttie substance from the part:icle.
1.0057:1 Where a range of values is provided, it is iixiderstÃ?od that eacli intervening v<ilue, to ttie tcÃit}i of the tjÃiit cyl' ÃbC lower IiÃviit Ã.MlesS the context clearly dictates other.:vise, between the ~ipper and lo-~vcr fa~iiit of that ra.rig~ and any ot~~er stated or iÃiten,cning value in that stated range is encompassed witliin the invention.
Tlie ÃrF~-per atid 1ower liiiiit: of these SÃ~ialler raiiges, wlaich may iiidep~iideiit:ly be included iri ttie snialler rariges, is also encompassed wit&i:iÃa the :iÃivent:ion, subject to ariy specifically excluded limit in the stated range. Where the stated range includes orte or both of the liniits; ratiges Ã;\c:luding either both of tliose iiic.luded Iiinits arÃ;
also incltid.ed in the i ~iveiiÃion.
10088] Uii1ess defined othet-w-ise, all technical and scieÃitifte ternis Lised hereiii have the same meaning as comtiioÃaly understood by. one of ordinall= skill in the art to whic:li this inventioi} belongs. Although any methods anci materials similar or ecluivalerit to tliose described fierciii can als~~ be LÃscel in the practice or testing of tIle preseiit inventiozi, ttie preferred methods and materials are now deSc:rÃbed.
All pUb(ic;atior~s mentioa-ied herein are incorporated hcrÃ;ir~ by reference to disclose ~i-ici described the met1iods andiÃ.~r matci'ia[s in connection witli wliic.li thc pÃÃblicatialis are clt:ed.
10089] lt must be noted that a.s Lased, herein and in the appended c:lairris, t.llc singular forms .<a', .zagid"> and "tlae" inc(ucle p(ara1 referearc.e;; Linlesw t1ic context clearIv dictates othe~~,vise. All techniual cmd sciei-it.ific tc::ri7is Ãi.:aed herein have the sarne Ãrl eaninu;.
100901 The publications discussed het-ei~~ are provided solely for theÃr discloswre piior to the filing date of the present applacataoÃa. Notfiii-i4,, here:i11 is to be construed as an aclmiss:iozi that the p:res~rià :iÃiveÃitioii is iiot entitled to antedate such publication by virtue of prior invention. Fua~ther; the dates of publication provided inay be different froti7 the actual priblicatioÃi dates which may need to be irideperideritly corllirmed.
EXAMPLES
10091_1 The follo'wing c;xaniples are ptit forth to provide those of ordinary skill in t:tie art avÃth a complete disclosLzt'e and d~script:ioii of ~~ow to make and Lise ttie preseiit i~~~~enÃion, ~iid are Ãiot iiiteÃided to liniit the scope ol:'what. the inventors regard as their i~ive tition nor are they, intoÃided to repre;;ent that the experiments bÃ;1aw are all or t1le only exl:rerimcnts pcrtormeci. Efforts I~ave been made to ensiir~ accuracy with respect to numbers used (e.g. amz ~a.Ãnts, temperature, etc.) but some experimental errors and ~~eviat:ions :~hoL~ld be accounted for, Unless indicated otherwise, parts are parts by weight, ancs(Ã;cula.r weight is weight average molecular weight, temperature is in degrees Centigrade, and pressLwe is at or r~ear atiiiospheric.
E-varirpk- I. Tabric:irtio~n r,+af.thc C.:orataM~~~s (Particles) 1.0092.1 Fi ;uae i is a schematic dia~;;raan of the process flow used to fabricate tlle ~ C3 containers of the present inyrt.ntic3t7.1110931 First, a 5pAm thick sacrificial laver of poIymetlivl methaerylaÃc. (1'MNtA, MW=996 K) Nvas speiÃi on a silicon substrate. The term "spinning" as i.ised liereil) refers to a process whereby a fltÃid is dropped on a rotating substratc. A '1 ? in-n laver of chrtarnium (Cr) and a 100 nii-i thick layer of copper Wt~) were evaporated on top of the PMMA coated wafer. 'rhe ('r layer functions as an adhesive promoter while tile Cu layer fiiiictiotis as a cotiductive seed laver for subsequent electrodeposition. Since it is necessary to etch tl-ic Cr and Cti later in t.lic process, it is nec:.-es&iry to minimize t}icli` thickness to achieve a i`apid etÃ;(a.. l-Iowea=er, c) minimize the electrical resistance of the film across the wafer dLrrÃ.raig eleeÃ.rodepositÃon, tlae rzaaterial itaickraess has to be increased. A thickness of 125 tir-ii was cieei-a7erl optitnal fior the prewnt application.
After t}iiri film Ãiepositiora, the sLibstr-ate was patter-raed using pI-rotofiihÃ?gral,lay. 'f .he photoresist Shipley SPR220 (Rolarri and Haas, www.rolarialaias.coni;l was first spun (ara the wafer substrate, the thickness ~~f the p(aotcaresist was ccaratrolled by changing the spiti. speed aiid the titarzaber of coats. After a soft bake, the resist was exposed to tJV
light using a aiaask: alig.raer. `I'1ae photo.mask used to pat-terta the resisà was a transparency mask witli six 200 Ãarxt squares spaced 20 pm apart. After exposure, the wafer was developed and tlic thickness ol'the resist was xneasuredusing an Alpba-Step p.r=rs#ilomete.a=. 'I'lieai; e[ectrc3deposit7c3ri was used to biai_ld pattern ttae metallic faces of tfic container in the photoresist Ãiiofd up to a taeio;ht oi' 7r15 p.m, usir~~v; eortlftxerci~l elect:rolytic solut:ioais ("I~eclanic: Iaxc: Nvww.teclanic,corai) containing the metal ions of c(ioic;e. (:`u was electrodeposited -followed by a -kliaii layer (about l. pm) of gold (Au) to fo.a=ri7 r7oaa-~~iagaactic coraiait-iers at-id a tliiÃi laver- (abOr.It 1 pi-ail) of nicJ<el (Ni) to fabricate magnetic cc4r7tainers. The Atr was Lased, to protect the CLr sL7r~acr~ from subsequent etr;liiiag steps witl r:Ã;riÃlÃ;r it itiert.
J0it941 A secorid round of ~.,hoÃo[ithogr-a.pl~~~ Nvas performed in order to pattern the hinges. A second layer of ;ST'.R200 was spLira on t.lae substrate and a hinge photonlask was rÃsed to pattern the hinges. The hinge mask consisted of two 1;Ãnwis of'hÃr~ges (50 x 160 (a.tni"aÃaÃl 25 x 160 pna). The wider hsnges were at the interfaces of ad~lacerat fa.cez w l~ile the narrower }a~r~-es were at the ed~;es of tlie taces. AligrarneÃrt marks were used to ensure perfect aligrimerit of tlie 1~ingles to the faces of the 2D
pr~ectrrsor=. Prior to hiÃagÃ,~ elcctrociepoiitiorr, the exposed t_~`tr arrcl Cr in the area of the hlÃages were e c:heci rasiiag commercial etclaarats (APS-100 for Cu arad CRE-473 for Cr, 'I'ec}ariic, IÃie>
~~~~~iv.teÃ:.hnic.Ã:..Ã-araa). Alt[ioLagla tlae etchants 1iave a high selectivity of Cu or Cr ~~,ith respect to Ni or Atr, the etch tlri-ac was c}ptimizc.d, to mirrirniz~ ~~~~~age to the Ni or Cu/A.u frame of the c:~~~taiiier. Pa,tre; tlnl.m.p. 232 -`Qor tira/(Ã;ar.l 1Sn/Pb: na,t.~. 183 'C3 solder was then electroplated in the hinge regioris. The lreiglit of the hinges was approxiiiiate[y 5Lim to about. I -5 pr.a-i depending on tlic face pattern aDd ilie type cYl metal Laaeci (wettinM, or iioÃircweÃting). .A_#:ter electa=odep sition, the origirial seed laver was etched ~iid the 2D precursor ten7plaie was itt~tnersed iri asolution of N-Methyl Py.a=.a=ol:icio.aic (N-MI', which dissc~~~~es, the sacrificial PMMA la;:erw) to release the precursors fro:rtti the wafer. Approximately 50 precursors then were scattered in a sinal( ca-vstallizin4,~ disli LasiÃig a pipette. A very ii~in laver of RMA-2 flux, (lzidiLim Corporation, www.irzdium.Ã:,oÃrt, used tt) dissolve any oxide it7rttted c~ii the solder) lvas p(yured into tE-ie diSli. The dish was theÃi heated to 100 'C t'or abotit ?Ãrliri to about 3 mfn and tlieÃi rainped rgp tc) about `.2'50 'C t~~ about-300 'C for 20 SeÃ:,onds. BeeaÃise of the low voltiÃaie of fltix, the agitaticin, was sufficxettt to correct for defects in the fiolding but iiot large eriotigb to cat~se, the crosses to ct)(lide iÃit:e~ each ot:lier atid becoi~~~ fi-iserl.
The n7olÃen solder generated ttie l=o.i-ce needed to fold the 2D l,recur~ors irito 3D
containers. 011 Ã:..ooliiig: ttie cotataiÃiers were perÃ~anciitly }ield to(;et.her by solid solder rti iiges.
1[)095] Dia.magnetic copper (Cu) containers were fab.ricated witli linear dii-iieÃiSioÃis of about 200 pli-a (rvhcrL one picometer isff'M ii-aeterj. As compared to smaller or lar.p,er sized biocapsules, the 200 pm size provides tlic maxi~~~~~~
crÃcapsula.tion volumc while still allowiÃig the ditTtiSiott of oxy<deÃ~ and nutrients to the cells.
It is kt~ow-11 that if cells are more than about 150 liÃri to a1~oLit 2100 ~tm away frortl the nearest blood vessel, the environment becomes hypoxic rR.H. Thc3n-iliiison wid L:.R Gray~
Brit. J.
C'anc:c;r. Dec. 9, 539 ( 1t355)i. In princip(e, the fabrication strategy desci ibed hereiii also would work oÃi. smaller or larg,-er size scales in. the design of containerS
for otIier a.p}:slications. The 1Ãnea:r diitaenGioii of the container was orderS of magnitude t~-naller than the wavelength of the oscillating ~~agnÃ;tic field at 500 MH-z, which is the hÃ~~heSt operating frequency in our magnetic rosoriag7cc (ANI-R) scariiiers, Het1ce:
ttie size of the perforatiotis oti ttie faces of ttie container }iacl no cietrinientaI effect on the shielding characteristics of the container. The thickness of flie ~aces of the container was clc;.signc;d to be larger thati the conductor skiri de:ptli at the frÃ;quÃ;ncy ot'th<; racliation.
T}ie term "skin depth" refers to a measure of tlie average depth of penetratic~i-i of an c.lec.tÃ'omtzgnc:tic: ti.eld.irato a rnat.eÃ-i.a(. lt. is defined as i.he depth at whic;li the priniary electromagnetic (EM) t:Ãeld is attenuated byrclcc,re:lses to (lle) of the field at the sÃar#"ace, or to appi-rxxi~~iatciy 37 f.~ of it, -vaiue at t:i-ie scÃrf"a.cc of the shield (A.
'1'sai.iovtch; I:Iectromatgltefic JVtr=e1esa .,41)1.Ãlicctfions (Muwer Acatlertiiic Publishers, M.A, I ~99"),)_ A tl7:icker container also has lov~er conductor resistance, ensa.ÃrÃng t(iat the eddy currents persist long enough to maintain shielding during the time of image acclttisiÃzoÃi, I'lic sk.iÃ~ ~eptli of Cu at 500 M:E~-lz is about 2.9 ptt-i (C. Kittel, Intr(xlaretton (Wiley, New York, ed,, at 7(1995 )): }ieÃtce, containers were designed to ~~ve, frames with thickness ranging from about 7 pÃnto about 1.5 pm.
1[)096] :[terrornagnetic Ã-iickel (N:i} containers in aclditioti to the cliarnagneÃic t'l-Ã
containers described above were fabricated to investigate tlic effect of mag.ti~
susceptibility ori the MR images of the contaiÃaer. Map-netic field distcÃrtions laut nat IiÃiiited to, shape, aiiiplit~ide az~d phase clisioÃ-iiotis, resulting from the cliffererÃces in inagtÃetic susceptibility betvkeen an ob~-jeci and its surrouncling Ã-iiec~ium cause a loss of phase coherence in the magnetization of the sample. Since the magnetic susceptibility ot'Cti is comparable to that of -~,vateÃ~, while tliat of Ni is orders of mallnitude higher than that of waÃeÃ-, a more pron~.~Ãunced distcartitÃÃ1 was expected for Ni containers in ~queoLas media (L.W. Bartels, et aI., J. l EUc. lntei-v=
licOiol. 1 ~~: _3 )65 (2001)).
100971 fi'he strateg~., tise.d to fabricate both the. Cri and. Ni containers ià Ãvralved the auto-folcling, of 2L) metallic precursors using capillary forces. "Capillary ac:tion''', capillarity ' or c:apÃllaR, motion, wliicll are used interc~han1geably lierein to refer to the abilitv of a narrow ttzl,~e to draw a liquid upwards ~.~.~ainst the force of gravity, occurs when the adhesivc iÃitcmiolecular forces between tEic ligL-id aricl a solid are stralt.;cr t1iaai the cohesive interrnolecula:r forces within the liquid. '-['he same effect is what cauSr S porous materials to 5oak. L7p liclt7icls. Previous demonstrations of acÃto-lolding include the actuation of rÃiicroziicter size t.ompoÃaeÃats and the assembly of ~}D c~niplex stnÃc:turtas (E. Smtala,et al, S'r:ieÃrce 268: 1735 0995)7 P.W. Brec.ti. et al., J.
<~~r~:rcr- lc~~~tr~r3rrrt~~:lr. .st 4: 170 (I.~1~~:~) K.F. Harsh ~.t~ :;~l.R
At'ftÃ~tfÃ~f.>~~ .4 ~: '?:>7 (1 E399)l;_.. [_:. Htii et a1., If;t~.fsl .a' ~`} IÃrt. [.YrartjÃ; On X-fict'o .f.kc:ir'c~ Nfec:hcrÃfiec:zl t~.xttts, 602 (2000), D. I=4 (Ft-aeias, ei: al Adv. A-iÃrler . -l 4 ; 23 '? (2002)).
[Ut~98J .'4ccni-dirig to oiic aspect of the preseiii i~~~~erition, _>D. I-ioIIow, peÃ-)`ora[ed containers were fabricated from ~.D precursors. 'riie process used to fabricate the 243 precursors, which is an extension of the process described in Exa.inple 1, and reqÃiired severzil. additive lavcrs, two photolithex;ra.py steps, tNvo electzodepositikiti steps, and a.
precise sequence of subiracÃ.ive processes. Briefly, the process i~ivo_lved i3ati:criiirig the metallic 27D faces rflsi~~g pbotolithography and electrodeposition on top of a sacri.ficiai Iaver. The versatiiitv of the strateg y was demonstrated bv fabricating precursors wNinse faces c,orrt:aiiied two different patterÃis-----c~~~e pattern comprised a square ti-ar~~e with open faces, while t}ie otl~er consisted of a m:icroscale cross shaped patterii iti the center of each face. In asecoiid laver of pIiotoresist, hinges patterned on the edges of the frames. `I'Iie width oi'the lai~ige between t-~~o a.djacezit. -t.'aces wa.s t~-vice ttie widÃlr of the Niinge at tl-ie edges so tl-iat aCl Iiirigcci ,joia-iÃs I-iad equal solder volume i:~pw~
foidin9: the solder vo1~ime was critical to eaast7re a folding angle of 90' (R.R.A. Syms, et al...r .I. I'?; 387 (2003)). After the 1iÃziges were t.,a.tterried, the 213 precttrsors were lifted cff tt-ic wafer by dissoltitiori of the sactiticial layer, '1`he containers avere sel.tmassembled by 1~eating the precursors abc~ve, the rnelting point of the solder, wherein ti-ic liqa,tid solder with bigh surface teiision generated t1l.e :i=or-oe rc:c;tiired to f-o(ci ad~ja-cc:nt faces of the precursor.
1.00991 Fi,,g. 2A sbows aii optical ima<we of a c.oilect.icari of containers that were fabricated tising tt7e process outlined above. The fabrication ttrategy allows a large trUmb~,'r of containers to be constructed in a single process riin. The primary yield-linziting factor was the error in estiiiiatirig tiic voltitiic of the solder to be electr~~~eposited at eacb iiiiige. The spaciiag between t~~e adjacent taces was also critical-ysrhen the gap between faces was either too I'''.argx or when the faces were ft~sed, the yield of folding was greatly $iniited. Fig. 2B=22D show optical and SEM
imagges ot'the mit;respattemed containers at different stages of the fabricatÃon process:
the 21) pr-ecÃir5oi- with e1eÃ;trodeposited fiiceti, ilic preÃ;urtior witli i.~tc:es arid h:ir~geS, aTici t (-ie folded container.
1001001 Although a~~ opeti-Caced c.oa7taiticr B, iaot ideal for= ari enc.ipsuilation device, since it is corisiderablyleaky, opcn-1'aced containers were filled for easy visualizatiO11 of their cont:entS. For M vivo applicatioris it .rtiay be de5irabIe to use the described strategy to construct containers wiili selectivelv sea(ed or micro/nano perfcyrated faces, and :(=a:1?ricate r~iore c.oztiplcti; polylicdral containers with r-oLi.rtded vertices. A.11 open-faced container (Fig. 3A) was loaded ~Nvith rnic.r=o"bea.ds since niany cellular dcliaeo., techniques use microbeads witti cells adhered to t.faeir surtace. In order to load the corrtainer with microbeads, a susperision of tl-w beads irt ethatio1 ,.va=
pipetted orito the car7iaitie.r. '1':1-le suspension entered tl-ie container as a r-csult of capill~~~~ forces. When tft.c ethanol evaporated, the beads were lield together by weak van der Waals forces 4at~e~~ir~~ the weak intermolecular Rorces that arise frorxi the t:ra:risieiit polarization of a giveit. rtiolcctile ixito a dipole) (Fig. 3:B); the glass beads cnÃ.rid (ae released by agptatiorl of t Nic cor7 ta'a tie.r.
1.001011 In order to dcnionstrate cellular encapsulation, MD.~ A,1.13-23t breast cancer cel:ls in an extracull.trlar= matrix (EC"N't) suspÃ;i-asiori at 4 'C, were ~~~~~ied in t1w contairiers (Fisn). 3Q, As tzSed 1-rcreitt, the terrii extracelltt[ar rt~atrix refers to the complex staÃrÃ:.tÃrra1 entity sÃÃrroi-rrid~~ig aaid, sÃrpportirig cells that a:re touzid wÃtliiri mamma1ian, tissues, as well as one or more of its constituciits incltrwiÃng, bÃ7t not IÃmiteci to, collagen, elastin, fibronectin arid Iwninin. MDA-4\4Ba22s1 cells are representative of rapidl'y proliferating cells and irximortalizcd cells, sLrch as P'I'O
cells, used in diabetes therapy, and stem cells used M rerwenerratiorr. On incubation at 37 'C for 5 miai, the ECM sÃrStrenaic3n gellÃ;d: the cells were retained in the biocontainer= and c:or.rld be released by immersing tttc corita-incr in wairii cell ct.rltÃirc mcdiÃrrn (Fig. 3D), It was also possible to load the biocontainers witli a celI-ln,C:N-1 suspension withiÃi an agarc?s~
~avity. lrr this casc:, a suspensiori of 5% agarose gel was tirst micropipetted ('60 prn tip) into the c:ontrziticr trsi~g a stcrcotac:tic: manipulator. The gel adhered ta,~ the sides tif t}ie container thereby 5ea(ing the faces ancl Icai>iÃi~,= a void in the center of t1x container. 'J`hc cell-l:C"M strsperisiozt was t.lrt:.ra rngcroirrjectcd into this void, w}ricli w-as then sealed with a microdrop of awY.t.roSe M~~~ci.
1001021 To demonsrat.e that ttie c.ell, were viable in the bio-ccamaincr arld orl release, the cells ~~~>crc stained with the fluorescerYt: dyc, C'a(ceir-i :'tM
(Sigr-fia-Aldrich), which stains positively for live cells. Fig. :'IE sltmvS calcein-staÃr-ied cells witltiri ttie biocontainer and Fig. -3F s}iows release of live cclls from the container on immersÃozt in warii7 cell cÃi1ttare ÃiicdiÃarti. 'I'lte frames of the biocoriÃaiziers t-tsed iri tliis dcrnotistr-ation kiad a tkiiÃ-i gold or platiritirYi coating orr the interior faces i=br biocoÃ~~~atilailrÃy, qiric.e gold ~iid platinum are, inert or tÃnreactivc materials. Pure tilt aitd tMr`Iea.d based solders were used to fold the cotitaixrcrs. It may be necessary to use ot:l7cr solders cnntainirro; iricr-t lncta(s Sucli as silver arid 4old for errliarrced biocompatilaility. :(t: is also possible to increase ttie biocompat:ibility of tlie containers, bv coatirtg, the entire folded container witlt a layer of an inert metal (by electrodeposition) or with polytiier5 (by ianrgtersion or vapor coaÃirig).
1001031 -NnnHinvasivc dctcci:ion ol'the c.oriiainers was dcri-iotist:ratcd by cr'Ylbecldirlg the containers in 5% agarose gel aÃid imaging them with tiIRI in a 500 X1.Hz vertical btire Brttker Avance micr+,3irnas~i~~ ~ystz.m. For tho images Shm? ir Irc rc, a. s3D FLASH
ScqtrciiGC Ivith tlre cch~.~ tilne (TE) irt the range of 4-6 ms; a repetitiorl time (TR) of 50 ms, flip-angle of -10', and a spatial resolution of 25 P111 X 215 ~ÃÃn X 20-lun was rised.
The containers also were i~~~agetl using a standard spin echo wetluetice (meaning a pttlse> seclttcrice tiScci in magnetic re.soirancc irraagin<.~ based oti the detection of a st.=ain. or Hahn echo, w}iicla uses 90' radio.frequeaxcy pa.iIses to excite the ÃnagnetiSm and ozte or more 180' pt.Ã1ses to retocus the spiitS to generate signal echoes named "spin eeboeS), with sianilÃtr reaEÃ(ts. Fig. 4 shows MR images of a 900 }tm cli<t~-noter c:at.~sill~ry corrtairiirzg a CLi (Fig. 4A) and a Ni (fig. 4B) container embedded in agarose .;cl. A
characteristic signature was ol-iSen~ed for both ttie Cu a:ttd the Ni corttainers--tltere is a pronounced darkness in the region of each container. These hypointense (dar~.) signatures 1~ave bcc.i} observed before in N/1-Rl of laq4cr centimeter scale metallic coils (A. Shcnhav. H. Azhari, 3V1cr~,Fw. Rras-on. 52: 1465 (2004)). While the region of hypoiiaÃc.ÃÃsity (darkr.ae5a) in the .14IR iÃ-iizÃ~,se was coÃnpar4able -to the size of the Ãa.oTi-raaag.aaei:ic Or container, it ~,vas rnucla: larger for the feÃ=.rornzagnct:ic Ni contairaer due to a pronounced s-uscc:[Aibi1it~~ effect (L.H. Bc:ÃÃÃaett., et aI.,.l. AI)I)1.
l'h4:s. 79: 4712 (1996).
BA. Schueler, et al., .1: Xlcigrr. Resrarr, knq~,i~~~g, 9: 596 (1999)). '1 fae iÃYaages of containers :Ãraadc of a ,aveia material were similar for botli opeaa faced c.ontairaerS as well as cross taceci containers, shou;iiag t}aat the pattern of the faces had fittle bearing oza the MR si graaÃure at ttzi s size scalc.
fUO 1.041 RF shielding w.a.s simulated in a non-magnetic coaitaiÃ-ier with a {-:iaÃite eleÃ~~ent model for a 200 Sp scatc wire t`raÃxac that was excited by a linear polarized electÃ-orÃra4~netic wave. Fig. 4C-4D are simulation ro-SÃilts showing rÃiagnetic field d1st:o.a=t:innS in the vicita.itv of the cotataiÃ-ie.r :Ã.tad reduced field ma-aait-ude in the irtaÃcri(ar of the coÃlt ai t1 er.
() 1055 ] For maziv biomedical applications it is Ãaecessary toÃaon-ÃÃavasive:ly track an eÃicza.pSÃ.alatioÃi devic.e. 'I'lic (:Ã:a container of the present invention could be t-r-acl;:ed spatially aÃid teÃ-iip~.~rally with in flow thÃ=oia"tl aÃ-i S-shaped 500 }sÃ-ia diameter fluidic c.}iaÃiiicl_ The chaainel was fabricated by molding poly di~nethyl siioxane (PT~NIS) in an SU-8 pbotoresflst ÃiaoId that was patterned Ã.Ãtiing pt Ãotf-Ait1aography. The channel was seated with a. s~cc)Ãad, t'iat, oxygen plasma treated. PDNIS
la.yer, Polyethylene tubes were conaaecÃed to ttae inlet and otÃtJet ports of ttae cllaziÃie1, the channel was flEished with silicone c3i1, and the container wÃ:Ãs' inrrodiiced into the channel. Under prcwsure driveÃr flow, the container moved withira the clyanÃie1 andwaS
imaged at ditTererat positions; the secli-ieaxce of N~I:R[ images is Shcawra ira Fig. 5. This ready trackability wit~~ TNIRI. at a>erv short echo times, without the need for a contrast agent, highlig}its a ma~jor advantage of the 3D metaIlic: bioc:oniainers or the present invention as coniparcd to manly other erica.psLÃlati~.~n systems.
Exanip1e ,27, MMuMtican caf Near Alc~qtxetic H~.}l~~V in th~.~ rerg;ion r_?/`
tla~:f container:
1001061 To denaonstrate an RF `hicldirm effect, tbe near m.~gneti.c> field response in the iric..ilÃity Ã3t" t(ae coaztaiÃaeà was siztÃulat.eti usiÃa.g a fin.ite c.lcnient electroanergrÃet.ic siniultztion ptacktage, F11K{_} (EM Softwarc. & Sytitenis-SA Ltd., wwts.ic:ko.Infoi'). A
t:i-all-wavc i-iieiiiod of Ã-iioÃ-iicaits approach ~~ias used to simulate t:lae.taear mag-netic field iri t}le region of a 100 Eam wirc: fracrie wii}t wire segrnents of 8 tiiYa radius', as,'uanrng perfect electric conductors coated witli copper (coÃidÃac.tivitv ___ 5.8 13 x 10'7 `?.na`'). 'f .lae ,imulation of the cubical wire fiariie model was performed witli a linear polarized platae wave excitationat 500 ?1't~Hz; we used an excitaticati sotirce of i V/m incident oza the wire fraiia.e, with E in the z dizectioia aiid H in ttae v directioti (Fig. 4('). The copper wia-c fraria.c was assigned a relative permcability, of I. thereby yimulatiÃag only the RF shielding etTect and n~~t the susceptibility etfects. Fig. 4C shows the near magnetic fi.~eld a-espoÃ-ase in bot.ii the x-v aaid the y-z central planes.
1001071 l.aa cnnclusioÃa, the dcscrAbed sÃaate;,~~can be tased to flibric.za.ie 3D, arbitrarily trackable, biocorat~l ners that allow perfusion bctcveen the caÃatcnts of tlie biocotataiÃaer~ ~iid the si-irroÃiÃadiÃig medium.
'I'laese biocontainers are eracapstilaÃzozi devices that do iaot lose their detectability wlaezi loaded ~vitb biological coritcta.Ã. Due to their Stre.ta~;t:la and iiigii porosity; stacla m"eÃallic biocontaiiic~~~ axc: uscftÃi as basic eiet1~entS of a scatfE}ld to gt7idc the gror~~Ih of cells in _3C3. Since the fabrication strategy described. bcre is cora~patil_ale with conventional 2D
nzicrofa.bricaÃion, it also may be possible tÃ.~ add electromechanical modules for remote activation, -,vireless coraamLaiiicatioai: sigaaaJ processing, a:iad biosensing to the faces of the bicscotitai~iers,. to enable medical diag-nostics and therapeutics". The present invention also otivisioz}a that sucl~ ~'I%D cotitaiiie.rs, whicla fttracti~n as small Faraday cages, avill iiid Latility in z ~tiaer applications requiring electromagnetic s(aieldiÃig in small o>olumes.
E.~~~~~~e 3: Aficrqf4bt~ication c~~id Se?f-Avs~.anzblV qf 3d Aflcrobox~-y,v fi)r Biot~~edical AlyWcations ~r ~:1~?t=p~~t:~1 t r:~t~:
1Ã10:1.O81 The process t~sed to ttibricate the boxes consists of micrcrlabriÃ;at7on arid Sttr~ace tension driveia sclt-asscml.~Iy, [h.F. Harsh, V.M. Bright, & Y.C. I
ee. S~n.s.
Aciwricai=s A, vol. 77, 23 7-244, 1999 ; E..[.:. .[-lui, R. T. 1-;*owÃ,, & M.
S. Rodgers, in 13th .Irrt. C70.rf on pp. 602tt607; R.R.A. Syms, EM YeatÃnan, V'.M. Bi-iglii, & GM Whiiesides,,I vol.
12, pp. 387.417, ~'~(}0? 1 io fabricate atad fold a 2I3 precursor :into a 3:D
hollow ,Ãauct.ure. I"l~e fabrication process involved tfiree steps: (1) patterning the faces oti tlie 2D precursor (2) patternin(; solder hiiiges bemNreeta ttie faces, aild (3) selt=assembly of the 2D precursor (:(tig. 6) `I'be boxes self-assembled wtieii the precursors were heated above the meltiiig poiÃ-it of t}ie solder, where:in the liquid solder with high surfiace tension geiierated the force reqtgired to assemble adjacetxt sÃ.lrtaces. The fabrication strateg"y allows a large raainber of boxes to be constructed in a single process nin.
Copper (C:.`u) atid t-iic.kcl (NÃ) boxes have beeti fabricated with and without gold (Au) coated SurfiaceS (to it3crea~e baoiitettne"'a).
Defet:t Modes:
tOO1091 Several defect axiodes iverc observed (Ci4F. 7); hoo~~~ever tvhen the process was optimized yic:lÃi` as high as 90% froz~i a single wafer wer-e obt~Ã_irled.
Apa.rt f:rc.~rn obvioais defect modes Sucki as over electrodeposition that merges t:lle taccs.
;nmcnt cst'the Ia~~igcs i.vitb respect -to the f-iices, atid over or Ãs.i-ider etciiing of the sc;ed laver, the lar_gÃ;st defect gtc~ was ttae hei<:~~t of st~ld~:r`
Ã~l~:Ã;t~Ã~t~Ãi~.~~t~;;it~:d at the ~es. I:t: too Ãliticii of- too little solder is cieÃ;trodepoSited the structure OVer Or Li.rtider folds=. Ã11 order to deien-iiiiie opÃiiv~~~ira solder beaght ~'or 90"
.tiolding;. published design rules [R.R..A. Syms. C.M. Yea#.man. V.M. Briglat, & G.M. Whit:Ã.sidc.s, 3, <t~r~:rc~~~Ic~trr3r~rt ~:#rc~rric r.r1 ~~ ~., 'vol. 12, pp. :347-4:17, 200:~1 were ÃiSed. Additionally, iTi order to iiicrease the ei-ror toleraÃ-ice, liir~~~s were desi,,:ried t3etweeti adjacerit faces to be tzvice t(ic widtli of kateral solder re(;iot~s patterned along the edges of the faces.
During electrodeposition, due to the elevated temperature (200 : CY) the precursors ,rere agitated (due to convective tlow Ãn the flLaid in which the boxes were seIfwaSsembIed);
this agitation aided in correcting metastat~~e mininia (errors) aiid helped the box fold to the thermodynamic mirziÃ~iuÃii, L0atiiiw:
1001101 To demonstrate that the boxes could function as encapsulation deviÃ:es, they were loaded with a variety of medically relevant constituents inclt,tciÃ~~ gels, beads, (i.qÃaidsr and cells (Fig. 8). For easy Visiralization, boxes w-ith all opei1 tat.:..es wt;re.
tased. f=1owea=ei', .iÃa real applications, boxes witli oiily one c~~~eTt face fcir loading, w-iÃb the otkaer faces close(l or pi.grcÃLis-, wc3Ã71Ãi be Ã7;cd.
IÃ30.1.l11 The byclrogel PltarorÃic; F127 (201NO ;soltttion) e\hibits a thern-ioreversible transition from a l:iqÃa.ici solution at l~,,,v temperature (e.g. 4 "C} to an ordered micellar ctibic phase at roniÃi tetrzperattare_ This pi-operiy ra-iakes ii: vet)5 attractive in tttc storage and releast. iia datag cielgoe.ty. The 1Ãvtizoge( consisted ot':;t 20% w!"w rÃ-lixttare of1?ttaroDic.
I. l d7 l,pt:Yly(etlav1eiÃe oxide,)-blot:.k-lacaly (propylene oxide)block-poly(ethylene oxide) capolvzrÃer, (BASF C 'c~rp. -www.bas1'.t;om) in wat:er. ~l'';'tie saiaip_le was staakera tisila.4,) a vortexer to speedup the atixin.g pÃ`ocess and stored Ãtt 4 `'C be-tbÃ'e usage.
In order to load the hydrogel in the bo\, a drop of the liKltticl solution was placed sati the box. ;l7ta~
to the b;.droplailic: si~~~v~--alls t.~-1`tkie ri~~tal(ic boxq the solt.itit.~ta r-eadily eÃatered the box.
'E3t~s.a.es were also loaded witlà "~=1==;DA11!fl:B-'' 31 breast cancer cells era-ibedded ir1 exÃ.a-itcel:ltalar rnaÃ.t-.ix 41:C:1~1~ gel. (NV1:C3A-=k-1B-23 i cells a.rt-.
representative of rapidly proliferating or .iÃa-irÃ-Ãc3rtta:lired. cells such as fiTC'3 cells tased in diabetes therapy, faTid stern cells Lased in regeÃ-acrat:ioti}. FÃg;_ 8C shows: a box loaded wit:li cancer ~ell~ that were briefly suspended in ECM glel at 4 'C. 'I'lie suspextsic?tx was introduced into the lanx arÃd was kelat at 37 "C for l:5 r2aÃ.ta to allow the ECM gel to polyraa.erire. The cells were stable in ttie box, ;Ã.tid could be released (Fig. SD), by pulsatile agitation of the box. ':I'}aes~ experiments clemottst:rate that it is relatively stt-a.ightl<cyruard to load tla,e 1?o.x.~s wiÃli a variety of coristitLietiÃs.
lnterartion with RF Fiellds:
lÃ3O 1.121 Sirice tl.ae boxes ai`e rnet.aligc tl-iev iÃÃtei-at:.t witl-i :RF
fields and belifa-ve its Faraday cages. This fÃsa.ittrtr tta~ bet;n Ãtsetl Lo ctet~.~ct. Ãtrtd track.
the boxes rt.n.totelrr tÃ:;ino.
magnetic reswaztrÃce gnaaging (N'l:RI). A t}.aaÃacteristicsi<:ÃÃattare was sabserv~.acl fot bcitb the Cii and the ^+;:i boxes-there was a pronotanced darkness iTi the region of each box.
This hypointense siwt-lature t`ac.iliÃated .re-ady trackabiiity wit:li M:Ett., at siroÃ-t ecl-io tiriies., without itie need for a contrast agent, arid highlighted a major advantage of these encapsulation devices as eoÃ-irpared to existing polyme.r:ie systeais.
1001131 Since the boxes iriÃeract witlr RF fields, this firatLai-e su,= ests the possibility of inductively hea:ting the box using a remote RF field generated by passing azl aItezzrati~ii), ctirr-erit tkrre~uggh a coil [:E. J. W. `I'er Maten and I B.
M. N-Ielissen; vol. '28, nn. 2, pp. I 287-1 `?9i3, 1992; C. K. C'boLi, in -5r`!r C "orrf i=trmr=zrtrzentalroa,r trml <,WeasurfAmc?n/ Iechõ 1988; pp. 69a7 r, J S. Curran and A. N-1. Featherstone, J.. vol. 2, rio. -1: pp, 157-160, 1988; K. Hamad-4chifferlx. J.J. Schwartz, t.T Sa:zitos, S.
Zhang arid IXII. J~cobson, Ndirrric , vol. 41 5, pp. ! 321-15-5. 20021. 13oxes coiYiposed of dia.a~~~gnetic t.CLi, Au) and ferromag.Iletic (Ni) metalc were fiabri^ai:ed.
W~~~ii a box iS
placed in a coil throu,,,,,h wliicft aii AC curreat is passed, aii electromagnetic force is induced. According to Faraday and :L.:eriz-'s Law, E =-N" aIr` , (t ), where E
is eIectromotive fii}r~~~ (Ei~I:F) indueed :in the box, tp is Ã-nagrret:ic f'l~ix generated in theRF
coil. and N'is the number of the coil tums. The i'=.ndt7ced E:~~~1~' caLiscs a cLirTent to flow in the box which can cause Iie.atii-ag, TIic heat generated can be calculated a.41' = .~-~,'` R.
(2)Y. wlier'e 11 is the heating power ger~~rated by the cur-r-errts, aiid R is tlre resistance of t:1le sample.
1001141 The alternating current i-n the box is stibjec t to the skin-dept.~
~hono-nienon, i.c. tlic c::trr.re;nt den.si.tv decreases with deptli. Sirice the thickness of the surtkes of flw boxes can be controlled with a raii9c of tbicknesses limited only by the photolithographic as~ect ratio LÃsed to patt.em the 21D precursor, bka:~es may be fabricated with wall thickness con~para.b(e to ttie skiii depth to mitiimize the e1eotrical resisÃaire.c. Additionally, if the box is ferromkgg7etic (e.g, Ni}~ t.he. h eatirrg is iarcreased due to magnetic hysteresis. As the prima~y purpose of irrduction heating is to maximize the heat oncrgy generated in the box: the apert.tirc of the inductive heating coil is designed to be as small as possible wid the box needs to be fabricaied w~~~ith a matena1 that i'eattires low resistance and high pc;rnieabilitv.
1 (1{}l 151 '('wo kinds of configurations were d.eincmst.rated. [n s~iie case tl-ie btsxes were iritrntluced iiit~ a v:ial arouiid wliieli is wrappecl a e.Or~ coil th.roi.E4,~li which AC
current is passed (200 M'E4z tc) I C31-1:r, 0 i tcz I Watt). In order to itite-grate heating of t(-ic boxes wiili '?D Ixzic.rofluÃd.ics, and to maximire inductive coupling, 21:3 coils were also flabr.ieaÃed. "I'he 2I) coils ar~ fa~iicatecl photolitEio4>-rap}iically (Fi,x 9) aiid cari be made with a varieÃv of ta.irns a.iid s~acrng. 'l`he box is placed along, the central axis of the coil in order to maximize iÃiduetive heatirtg. A:liliougb the ziuÃiibez"
of turnsin the '?D coil is less tliarz tE-iat: o1'the 3D co:il, the eavÃti, of'the ?D co:il is eorriparable to the ..,ize ot-the box to maximize liiduetive coupling, hiduct:ive heatino characteristics of tlle bnNes aÃ-ld the coils are 1ieijag measured.
R.eleasiti; a chemical froni the bcax upon hÃ:atln ; ~;
1[)01161 :I.n order to demonstrate Ãhat a cliemicaJ cotald be released irnrII
t:l~e b nx-upoÃl k~eat1lig, ilie l?c~.a was loaded with a l7ydrr3gel that was dyed red.
lnyÃially, one g.a-ain (.>C l'(ti.a-ÃsiaiÃ:. F88 (Molecular weggftt.: 1. 1.400.meIting pÃ:aini.: 54 'C, obÃ_ainecl t:rom BASF) was c{issoived:ir.a 1.0 anl. of aÃ;elonc:, '('lle sainple o-vas t1~eTi heated aTid sonicated to aid disso[ui:inn. A few drops of the dye erythrosine were added to the sÃ?lutlori. Ari open fkiced box xvas loaded with. tlae dyed lavdroge1 solution usi:ti- a sy.rin~e and the box. was allowed to sit until the aÃ;etor~e evaporated. Sr~-icÃ; the hydrogel dissolves M
water, the loaded box was iÃiimersed in dÃ.~dec,iaie (hydroc'el does Ãiot dissolve i11 dodecane) and placed on a glass sd.ide Tl~~ slide was heated tÃ, 70 'C' on a hot plate and optical. photographs were taketi at 3, 7 and 10 m.ii3ÃitÃ:s. 'l`lae ge1 softened t~iid the dve was released icito the dodecane solution ffig. .10). The release of chemicals fircam other l7Nrdre~.o,els in water, as ~~~ell as c~l~tirvliri~i~; inductive RF
t~eati~~u~ of the loaded boxes, is Ã:.Lirr~iitly being ~~ivestigated. For in-vivo applicatioxis it will likely be necessary to Eiea:t tlie box approxirnateiy 10 "C' above the temperature of the human bÃ`~dy .
CONC: LU~SiONS
1001171 Tia starni-nary, a azew erac.ap sÃa.IÃttic.iia device platform that cÃ.~iaibiriÃ;s ttrc tavtarab1e aspects of three dimensionality with Si microfabrication has beerr dÃ,'rt-itansÃrafed. Development of devices rrsiÃb 7aanopt3rous faces fclr cell encapsulation therapy (without imr-ntarac3stri~press.ion) and ciesiizÃairig boxes with optinlired RF freat.ira_g profiles for reriaote relea;~~ of chemicals as in prr.3g:ress, E.Ya~~~L)le 4: R~.pnirt~.t .~ Radio-lT~rUt u~,pn. c ? Controll~.ad Nano1her Ch~-yntistr v arrd Cizeitir.c=y l I)ef.,avc~.rr, (rn .S'y bgrat~.~s, 1[){1118] Containers }rave been tab.r:icated Or.rt Of ta-leta.i, wh:icli a_lioNved tl-iem to be rer-zirst:e1v coupled to electromagnetic sorarces. I'lY:iS ~eatrare was used to eraable wireless control over btrt.}a the spatial guidance (using magÃaefic containers) as well as the delivery of naraolitr,r vo(uziir,s of chemical reagents. `I'i~e containers cati be gr:iidrwd in spatial p,a:ttems tE-iat: are aiot lii-raited by flow proffles.iÃa eotivetatiorial aiierofluidics, that is, downstream fto:m a channel iiiiet. Tt7e remote-controIled iiaric?liter containers enha:ncet~e capabilities of present-day xAcÃ-oflÃrid:ics by enabling spatially cÃ~iitrolted c(icziiica1 ~~eactioÃiS, miÃ:rofiabricakiian N.~iiÃbin capillaries, aald ola-dezYiatid localized de1iverd~ of chemicals to crxrlÃu.red ce(ls_ ~
100119_1 A combination of conventional mier~~fibriÃ:,atiran and self-assembly [1-'. G.
f,..eorrg: Z. Gu, T. Ko[i, D. H, Gracxas, J. Am, Ã;laem. 4oc. 2006, 128:
113"36-11 337; &
CJimi, T. LeoÃag, Z. Grxa, M. Yang, D_ AÃ-teÃiiov, I. N-1. L3tlr.riwa11a, D.
H. Gracias, Biomed. Microdevices 2005, 7, 341-34,57] were used tÃ) fabricate g0ld-coated, nickel naÃaoliier contairicr5 (FigLrrÃ; 1I a). To facilitate chemica1 delivÃ;ry, the c..orltairier-s were filled with a gel that was soaked in ti-re chemical reagent to be released (Fi.;tare 11 b).
'B.N~o gels were r_.rsed~ pluroraic.151 for general dry-release exprwr-iÃnentS
and po1y(Ne isopr~~py1acry1amide) (P1~IPAna) [T. Hirokawa, T. Tanaka, J. Cheni. Phys.
1984, 81, 63i9-6;>80; N7:. E lsl.am. A. INI6~~saved, Z. DogiÃ. J. Zbang. T. C.
Luberi41cy, A. G
YocJii. Phys, Rev. Lett. 2004, 9`21. 088303] for cheriaicai delivery in aqÃreoÃrs solutions aiid to living cells. P1r.aroÃaic is a water-soluble block copoIvraier bydrogel that scatteras at 52''C and is c:ompatiblc witl-i aiv.idc ran;Fe of chemicals [R
A1exa.ndridis, `I`. A.
Hatton, C:`oiloid SLart. ~i-ivsicoclieÃii. l;no;. Aspect. 1995, 96, 1-46].
Hydr-ogels based on PNIPAmIfi are thermoresprxrisive materials tliat are widely crsed in drLÃg delivery, because Ã1~ev ut-idcr~~o a structural tra.rasÃtion racar- the temperature rai-r,,~ of tl-re I-ILtmarti body [-H. Yu, D. W. Grainger, J. C'ontrolled. Release 1995, 34, ilittl?i; K S.
Soppimath, `i'. M. f"triiiriabliavi, A. M. Dave, S. G. Kurnbar, W. E. J-',LrdzÃraski, Drug Dev. Ir7d. Pba.rm. 2002, 28, 957-974]. This transition temperature, as well as the collapse kinetics ni":Ã'N1PAm, cati be altered by addirtg co-moticri-rers aild cliarigirig the degree of caoss-linkino.- [(~_ A. Stile.; W. R. Burghardt, K. E. Healy, Macromolecules 1999, 32, '7370-::379]_ Hence, PNIPArn i5 ati ideal candida:te for remÃaÃe-con trÃ?lled release to living cells at-id in liquid ri-iedÃ'a.
100 1.201 Once loaded, a container was placed iri tlic react.ion vessel of elioice arid could. be guided in ariv spatial tr,~jectory Ã.isi~ig a magnetic stylus. After guidaÃice to the desired 1oca.t:iori, a radio-firequericyr (RF) #Ãcld gcrierated by a 243 micÃ-ocoil wa.s directed towards t:lic coriiait-ie.r_ The pov,~er in the RIF field coupled iriduc[iaelN? Ã:o the metallzc container, thereby proclLrcing eddy cLÃrrcnts in the frame and heating it up by a Joule effect. It is possible to heat c;vÃ;i-a ~onma.gnetic metallic containers by indtictivc c.otiplirz.g, ari~.~l tlie heating meclia:riisrri is clitTererrt from tl`rat.
used to heat ~.~Ã~~lymeric magrietic microspberes. ;Sirice the coratairxers were inic.rnfibdÃ:,atcd, the electr'rcal characteristics cou(d be made reproÃiÃrc.ibIe, and the temperature coÃ71c1 be precisely controlled by changing t1ie irZciderit power. This rc;prodticibility shou(d be cÃ,`~l rtxa.sted with t1~e pozver needed for release trorrx polymeric magrretic microspheres, wliich cazi vary greatly ~ecatÃse of pol~'dispcrsiviN, in sizes and inhomogeneous distribution of magnetic particles within clrffererit macrosl?hc res.
1001211 Blv heating thc corita:incr, the gel encapsulated ztiitl`riri it sotteried (crr collapsed) and released the c.bemical at tlic targreted spatial location ~Figure 1 Ic.i. `l'he metallic cc>mainers are essential to obtain heating at the power and frequency settings tisc.d. No r:clc.ase was observed from tlic gel in control experiments (on expa,~sLirc to thc.
R.F radiation, btià :iÃà tlic absence of the container) bccatrsc of ric{,~Ii~ibIe dielectric heaiin_g at tE-ie kÃcqucricy arid power sirtttÃ.irgs tÃscd (see the Stal,po.it:ing Iriiormati(~n).
1Ã101221 The rettrote-coritrolled c:oa7talirei-s, rna:14_e it pi5ssib1e- to do cherrris-tr~, witli unprecedented spatial cotitÃ-ol .in 1-aardrtc3ri-eac.li reiorrs. 'I'o hÃg;hfigtrt this feaÃtare, Vve repaired a break oap in o:ire of two aqjaceÃit microwires embedded within a capillary;
the capilla), was accessible orily by hipt.lt aiid ~~t.ttpttt ports (Figure 12), The gap witliizi microwire 1wa.s repaired by remotely gtaidirig cntitaiziers to that site iti air Wi~;i.irc 1"
a,b3 and reirioÃely releas:ing first a chemical scÃisitizei= atid tltei-i an activator (t.tsitrg two wparate containers) locally at the site of the gap (Figure 12c). "1'he sensitizer and activator were titi ai-ad palladium catalysts, respectively, which facilitated the electroless deposition of copper. After scrisita.r:itrg atid activating the spatial rcgiwl withiii the gap ofmicro-wire lorÃIy (t i,gure 2d), tlrc eiiÃ:irc ca.p.illaiy was flushed Nvit}Ã a commercial soltÃtiott of copper su(fate (1~igtÃ:re 2e), <'11ttioi.ig}i both i-rucrow.ires and tlie wa1ls of the capillary wiere exposed to the copper su(1=aie solution, rtieÃaffic copper deposited oifly at the c}icmÃc.ally scns:itized atid activated gap in t-iricrowii=e I (t- igurc 12 f.). Electrical resistivity measurements c:crtifirmcd electrical continuity ol'microwire tac:ross, the gap. This result demonstrates the a,ttfllity of the c(irit~nor=.w for 1crc:ali,zcd clierzrical delivery and chemistry within capillaries arrd ~.~tEier small spaces. (iI
comparison to alrea.dv e5;.istinO; Ãiietliods of Ãnlcr~~affibrication in capilla:ries. J. C.
?~=1cDonald, ~ M. W(ritesidÃ;s. Acc. (::lrem, R.cs. 2002, 35, 491-=199;
MiaclotÃ, Ft.tnclanic.Ãita(s of Microfabrication, CRC, New Yorls. 1997: 13. J. A. Kenis, R.
lst~~~gflov, G. M. Whitesides: ;ScieÃicc 1999, ?85, 83a85]', this itxventioWs ziiethod is Ãiot limited bv . the geometry of tlie c:al,iIlaai= or laminar flow profileS.
1001231 ~secoÃic! demtÃnstÃ'atiotr. highlights the titility, o1't.tre rÃarÃo(iic.r containers in the r-cizioÃe-con t-rollecl, localized dirlivcry of sub-nanoliter volumes of chciiiicalw to specitic cells cttltuaed ori strl-istrates, Containers were loaded with I'NIPAÃn, soaked in a liv~.~:dead ('gaecnr`rcd) two-color fltÃcsresc.eticc viability stain [Irt-vÃt.t-ogen liv(-Aead stain product giiide to stalÃi cells 1ocally in a crjlttÃri dish aircl to t<t.~rify that Ãio Ãiet;i-i_stic cell death cscctÃrrc;cl dLariÃau cltet-nical release as a consequence of the heat.iÃÃws [S. Cc3r-On, S. Boexcb, C'. Maneschg, Ã.'. RacirÃ-layr, G.
.(~arfsc.~~, R
Klocker, ELar. Uro1. 20430, ;' ) 7, 499-5,04] or exposure to the R-E
Ã'a.diatit-11.t1_ 1001241 The L929 mouse fibroblast cells were cLÃ:(ture-d irt -35,-mtià well-plates wiih glass itilays and growrz to c.c~~ifltic:ÃÃcy. At the start of the rernotercreleas~ ex~erimcÃit, the ~prnwt:li media was removed and the cells were rinsed witli phosphate-buffered saJine to dilute tlie sertÃm esterase activity, tbere-by rriiriimizirig backg-round fluorescence. '~('o eÃiable reÃiiote release of the staiia, ati RF coil was placed below the plate directly tjÃider a coiitaitier, aÃid the coil was powered i.Ãp at '-2r3 W for tÃ-iiiÃltlt:e to collapse the. encapstÃlated PNIPAm aÃid refease the stain. Fliiorescent images were obtained 30-60 minutes after Ã-elease, toa.Iiowsufficient time for uptake of the stain. It is c.1eaa :i:rÃÃni the coiii"ocal fluorescence images (Figure 13) that the stairi was released loca11a<~.z~~~ ~~~ithiii a.Ã~.~.d.iuc c~l`-iess than 50~~ lt.i-Ã~. from the center c~#the c~~r~tal~~er. (t: cati ~
also be seeÃi that the cells exposed to the stain ha.d green fltÃcarescence, thtÃ;s indicating that they were alive, and Ãio red fluorescing or dead cells were obsened.
"('he results iÃidicat.e that neither the temperature used to collapse the encapsulated (''N:I:('':1m ri(ar the RF radiation c,a.used necrotic cell death. It should be noted that no lcakaye or spontaneous rÃ;1eas~. (that is, Ãio cell staining) was observed fÃ-uni the containers in experiments where Ã~~ RF field was a.pplied. Biocompatihility studies show Ã1o necrotic cell death occurs in the presence of the containers over 48 h.
1001251 Tn CcglIClLÃSitÃn, the metallic, self-assembled nanoIiic.r containers cwi. be titilized for rcniote>-coÃ}trol(ed rÃiic.rofabricatic~ti and cheniica1 cic:livery in hard to reach spaces. 'i'he containers will I~e useful in tabiicatiÃig complex and reconfigurable microarialvtacal, microflÃ.iidic, and micro-elecxromechartieai tvsterns. The localized rcnicitc: dc:1iven, of chemicals to cells establishes a methodology for r~.motely manipLÃlatiÃig thc chcmicai ajid biological micrv-eÃivirvnmerit for applications in cell engineering, tissue en4,~ineering; a:iid drug developÃiieÃit. Fina1Iv> the containers pro4.,ide aii attractive platform for the intogra.tioÃa of addit:ionai features of wireless devices (for emiÃnplc, frc.qÃ.eeÃic:v-sclective remote coÃitroI and remote coriÃmunication) with the delivery of tiancs(iitar volLÃmes of c.licmicals.
~XPER1MENT-AL 17E'll~,UL'-';
F'fkbr=iraÃic~~~ of the ilicroc0titainers:
1001.261 Brieflv, the fabrication process involved t.lre self asSemt7ly of a two dimeÃ-rsiorral (":D) template irrto the 3D cii.bic cnratainer. First, '?D
met.allic templates consisting of six square pc3r(yuS fiac.cs were pbÃ~to(ittlograpliica(ly patterned. A Secorrd layer of photolithography was used to pattern solder hinges ~ii the Ãxute:~-edges arid irI
beÃwcerr faces. `('lie '?D template spontaneously fotdeel iritÃa t.}re 3D
cubic contaiÃler wIreii it was }ieated 6rr a 17.Ãiiel) above the meltirig point of.'ttie solder liirlge5, wbereiri t(-ic surface tension of the ri-iolteri solder provided ttle 1`orce to drive Selircassemb1y.'f lrc final size a:trd porosity of t[ie 31) container was varied by patterning the 21) template alapro-pri.ately. In this experimerxt; containers were fabricated from nickel (Ni), a zrragrretic matcr-ial, to crrable rernotc tYuida:ncc. I'tie oiiter and ir-irrer surffices of the containers .:vcre coated witl-i gold (Au) to irac:rcasc E?iocoarpatib.ilit;:
ai-id decrease electrical resistance (low electrical reSistaiic.e, iracrcases t.lre skin depth for penetratiari of electrornagnÃ>tic: waves). The fabÃ-iUation process was highly parallel and large nLrml~~rs. of coritainers could be 1'a.bricated in a cost-effective manner.
PREPARATION OF TIJE C;ELS:
1'Ã301.271 PIr.rronic&: The gel ivas made by coa7rbirrirr : O.5 g of F68 Plrarcarrict (BASF) with 0.5 mI. of water. The mixture was sonicated for 10 mirrtF es to c2isurc complete mixing. Gelation c3c.cLirred after excess water evaporarecl.
1Ã101281 PNIPArii: Tl-ie Ã''Nl;Ã''Am gel was made I-roiir two stock solLitions, A and &
So1r.iiirYn A. consisted c3f.' I..6701. ~,s N-i.soprola?ilaca-ylarr-fide (1?Ni:l?Arrr), 0.0083 g N, 'ti'..
meth.vIerreb:Fsacr-ylaa:r}idÃ; (BIS), and 15 n-aI_, rarater. Solution B
consisted of 0.0129 ~ of amnionitini persult'ate (APS) and 15 mC. of water. Both solutions were vorte:~ed r.indl the solute d.i`solved. Gelation was <ac(i.leved t?t' mixing equal volurr-i~s of cacl~ soluficXi togetlrcr witlr 0.4 r~ (,v`v.) of Nv N, N`, N'yteta-aun~.~drvlenctbyl~.~rrediai-trirre (TMG[3) a.rrd oc.cr,trre(l w~ithin 5 niinErtÃ;s.
REMOTE dC-ti1DAN-(.`E;
1001291 Remote gtiiciaÃice was aclaaeved by inanipula:tir~~ a ~~~ag~ietic stylus below the reaction vesse(, ln ortter to redLÃ.ce t.tae f'ric.t.ioti between the comaÃnE~~~~ ~i-ici the su.r~ac.c of the vessÃ;1, the stylus was rotated alÃai1~,r the base of the vessel causing the Ã:Ãantainers to tumbl.e along the surface.
'i'HE 2D lf~U..N.11CROCt)IL SE'1'-UP:
[U0l.3()1 A ":D gniczoco.il was fabricated using phot:olithc~~~aptay on a f3r-irlted circuit boazd (PC~) as the I-',:F source. r1"be mici-ocoil was placed either below or above tlie contaiiiers at separation distances of approxiiitately 1-5 Ãnan. A c,lÃrrent at 800 MHz (RF) was passed t}irougli the coil to geiierate ar~ afteriiating zY~agÃletic., field iti a directiora perpendicular to the surflace of the c.oai; ata incident power ita ttae ran,)e of 1r7 Watts was usecl. The sufface of the coil was air cooled to remove any JoÃiIe heat gerteratec1Ãn the coil.
REMOTE R.EPAIR. OF A MICROWIRE WITHIN A CAPILLARY:
1.00131 1 M.icrow<i res (100 ~tm thick copper wi''.res, spaced 2 iiint apart) with break gaps of 50 ttm were fabricated ~~~i a glass slide using photolithography. Tlte capillary was formed usii}g po(yci.imethyI siloxane (RDTVt:S) walls that w~c;re scale:c~
against the glass slide (containing the wires) a:iad ariofliefl glass slide (the i=caot of ttae capi11ary ).
Seal'in4,~ was a.chie-ved bv plasma sa.Ãrface modi.f:ic:atioit of the :l'DMS}.
The capilia.t-y with the eanbc:cl.dc:d 3vire:s (f:=igÃarÃ; f?.A), wa.s approxiÃ-nazely l mm in widtla. and 1 .5 ci-n in length a.tid was ~.~taly accessible by the iiapLat a.nd ~.3titpuit ports it itti two eaids.
1001321 The sensitizer soaked E'la.Ãrotiic~gD gel was prepared by Ã:,ca.mbining 0.5 ztiL of sc;nait.izing soIÃiiic3a~ (Transene) witla 0.5 g of Plitronrc F68 (BASF). The activator soÃiked ) ~'1u.i Ãsni e ~ ~ gel was prepared by 7ng x,i.ta g 0.5 ml., of activating ;;c~l tati o~.ra (Transene) 0.5 g of pIar~.gnac F68. Prior to loading, eacii. mixttire was sonicated for 5 minÃites to enstire complete mixing. A 1 ~tL drop of eac.h of the mixtitre.s was pIaced on two separate Ã:ontai.riers ar.ad ttae solutions weÃ-c allowed to gel Ã.gvernight. (_14 hours). The containers were [bera cut out of the gel, to eiasca.r~.~ that the gel reniaiiaed only withiaa the cotit-ainer.
1 (1{}1331 After w*uid.ir.ag tI-ic coaztaiÃiers to the gap Mitbirl IÃUUr:arN~ire 1, WC ~t.iral applied 4 Wat-ts of power to tE-ie 2D coil until the gel softened and released the sensitizer solution from ilte container. T1Ãerà the power was t-edticed to _3.0 Watts -Lor l n-IiÃlLtt.e to allow sa.il='==:ti~cietit tizite for dift'us.ion Of tltC sOlÃ.atiOÃ-I through f1ic gel and to the surface ol the gap. 'I'he cl'aatine1 was tl'aeÃi flÃ.rs}iecl w:it}i water to reÃiiove the container aÃi~ excess geI. 'I`his process was then repeated, fk-.)r the secoÃid coritainer ti(led with the activaÃor solutzorl .
1001.341 .'4.ai electroless copper solution was Ã-iiade by rnixirig eomtiiercial solutions, PC clecÃreiless copper solution A arid PC eiectr~~less copper solirtion B
(both from ']'raÃiserÃe), irà equal voitÃriies. A .yz-int~e, wit.là a diameter of 0.9 mm was fitted with 0.9 znrri 11) ta:ibirig, 1'he (Ylkzer end of the tiibe was placed iri oÃ-ie openingof the channel. A
syzir7ge pLartip ((tAZF<.1.) was used to flow the platirig sc)Iution irito the channel aÃid over the 13rokeÃi microwires, A pulsatile flow was used to taciiitat.e the platirig reanon by maintaining a high local eoriceritratzori of copper ioÃis while <iilowirtg sufficient time t`nr deposition. During tE-ie e:x.perAiiaerat., the copper- plating solution was kept at.
45 C , REMOTE CONTROLLED DELIVERY TO LIV7.Nt. CELLS:
11101351 CoritaiÃiers werG loaded with PNIl'`'AÃti and allowed to sit overnigIlt, O1l the day of the cxpcr-ir.Ãient. the L.ive;/~~adO two-color fltrc3rescence stain (1:nvÃtrogen.) w:-Ãs prepared at concentrations of 0.5 cÃM Calcein AM aÃZd 1.0 ~iM EtlÃidiÃ,rrii hoÃnodiÃnor-l..
The PNIPAiÃi filled containers wei-c sa.ahr.a:ierg,,ed in tlzest.aiaÃ
st}lLrtion for 3.5 ilciLar-s prior to beginning the expei-iii:aerat to allow the PNIPAm r~t-n~le time C.k) rehydrate and absorb the liv(-~Idead stain.
10{ll361 :t.,9d~-~ mouse fibroblast cells (Sigma) w~,ere cultured and rrÃaiÃitairÃed tollowin;standaa=d c;ell. culture prc3toccais. The cells were cultured in 75 crn2 culture flask in 85% MiaiimEÃm Essential '~~Ã;ditrm Ek(gles containing L-<;~lutt.amÃne and sodir.rni bicarbonate with (0% laor;~~~ sera.trii mid s-Grpplementec1 witlà N1_EM non-Ã;sserltial ainirÃo acids and ;aÃ3ciitirÃi. pyÃ-taa=ate The cells Ã~iaiiÃtaiaied in ari incubator set to 37"C w.i-tii a. wa.ter-saturatec~ 5% C02 at--ai~.~sphere, 1_,929 cells were sul~sicultr7:red 2-3 times per week utilizing tr-yp;,in-1'11')TA arid seeding tEic Ãiew flask at a cieÃ-isity of =x 104 cells.?rn1;. 1'he seeding deÃ-isity was veritied by rerÃao-,>irig a sample of tl-re tiypsinized cells, stairring the cells Wittà t.Ã-y11aÃi b11ie aiÃd cÃsitia..- a bemacy-t.ometer to cociÃit tlÃe nr:irÃilaer of viable ce[ls.
1001371 :Kernote release experÃr-rietits were conducted in 35 rnrii well-plates witli glass inlays (I G1assabott.omsdislies inc.) for optinauni contoca1 microscopy.
Brietly, cells were seeded at a dezisiÃy of 250 tL of 2 x 105 cells:rÃi1:, directly orito the glass iÃ-flav <i.r7d allowed to recà .fioÃ~ 30 rYtin to prorYtoi:e adtlesiotÃ. 'i`wo rYii_: of g;Ã-owth ÃxÃedia was then added ajid the cells were irlctgbated for 48 hnurs to achieve a confluent monolaveÃ-.
1[){1138] After rerÃiote release, the cells were iÃ-iiaged usiny, a Carl Zeiss contbcat microscope. Br=ietl;;, the.Ãnicroscol~e was setup witb lasers and filt~rs, i~ecorni-fiended in the Live!Deacl,,x assay protocol. C;alceiri AM was excited at 488 Ãar~~ and ethidium horÃ-zodix~~er-i was excited at 543 nan. D-Ye up-ta:l5e was detected witli filter- eÃ-rbes of 1:31;' 505.5;3 )t] (for calce:iri in live cells) arÃd I.P 650 (for etlÃieliu-rÃ-r homodi.iiler iti dead cells).
CONTROL EXPERIMENTS
HEATING CHw'~RAf.:TERllS'li'ZCS<
1Ã10:1.391 To demonstrate control oa ei- heating of the nanoliter containers with tl-ic incident po3ver of tl-ifa magnetic field generator, a tc;mperatiive control experiment was condÃ.tct.ed, The sc:ttÃp of the experiment was similar to otir other RF
controlled release experirnents. TIw nrÃl_y di1l'er=erÃc~e was the p(aceÃ-nent of a Norirc:(teversible C)Nl:l:GA:I:,Al=;iEi.: Label (Ouiega `I 1:,-S series)urader- the nanoliter corÃ'[air7er so that it ccirel(l be heated witl-i good thermal contact. The i_ernlaeÃ`ai_ri.re of the CODÃrÃiTier sUÃ1~We cari be dedLÃc:ed color changes in the labels th,it occÃir at 38"C, 49'CY
60"C' arid 7VC
respectively. 'I'1-ic iiÃcideÃ-ià power c3t`Ãhe rnagrieÃ:ic field was iricr-eased u-rÃtil tlie specific label cliar34ed color (after Wai.t.ing .fo.r approximately .10 seconds).
1"'iw_ 14(a) shows the col.or- c:?ÃarÃ-~t:: in the Iabe1. oser` whicb the containeà w;Ãs lala~ed Fig.
14(b) is a plcÃt of the Ãc:.rnpertatÃarc measured b~:r t1~~.1~il~eis vs ihac iai~.~iehe~~t pÃ:g~n~c:r'. b~' c;l~at1;~irr~ t~hre ita.cident power, the heating could be precisely coaYtro_lhed. t;t slaoÃald be rroted t:liat the exact depends oia t-lre sh~eei#ie coa7taiiaer used and the experit-iieÃit (i.e. dry, or The color c(a.a.aau;e occurs on.[v Lirzcier the eorrtaiaier showiaY, tE-iat: the heafita4,~ is wet release, urrouÃadiÃag local. (B) A pIot of the temperature meastared usirag the color inclicat(yr s s label environment) vs the iracicleia:t RF power &Ilfri:I SI' A'1"lAi.: AND -NIATI:R-AI.T SEI..f:.C-1'lVITY OF REitlO'I'.I:s .It.I:si_EASI. FROM
TIiE CONTAINERS
1001401 To deÃrioÃrstrat.e tlic material arid spatial selectivities, the fo(lowiirg cc?ntrot expeririierrt was performed. 'I'wo eorrtaialers loaded wit:hr (''lurotridk:
4~;e1 (soaked ~.vith focsd coloring) were placed 3 ri-iraa a}iart: fro.in each other in a Petri disha. Aalotlier isolated piece c~fge1 (ia.ot eÃacapsÃ.ilated witbiax a ~~ontairrer) was also placed in the disla.
The Petri dis[i was aligned over a 2D xriierocoii at a distance of ? nirYr:
such Ãhrat in the plarae of tlie disla, the isolated piece of gel was aligned directly over the celater of tlle coil: one of the containers was aligned within the c:hrcumference of the coil bcÃt off'set from the cc:titeÃ~ by 300 ag-iel the second container was nri;;a(ignÃ;(i and lay outside the 21L) coil, ft'I-ie coil was hsc),wered up at 800 MHz. Only the gel wit.hila the container aligÃred avithrira the circumferetace of t(ae coil lieated LÃp aiid scafteraed at a power of 4.7 Watts. Even when the power was increased to 7 Watts, the isolatÃ;ci piece of gel placed in the region of highest field as well as the gel in the raiisa(ignÃ;d container remained unchanged. This coÃatrol experhrraeÃat demonstrates that the inclticti~~e beathÃ~g had }aigla spatial and material selectÃviN,. The ex~eriixaerat also shows that the metal used to tahsric;atc: the nanoliter cvirt-ainer is c ssetitial to enable the inductive he<atiÃag. It shoiald be noted that a Ãnar?ttetic matei'ial is g7ot Ã-eqtaired to fiacilit.at~
remote heating (the magraetic property is used morelv for spatial gÃiidance). Also demonstrated was the release from containers wihha iio Ni, i.e. c:oratiposÃ;d of copper / gold.
NO DIFFU'SIt)N OF CHEMICALS IN THE ABSENCE OF RF R.ND1AT.ION.
1001411 A control experiment was pt;.rtt}rmed to clem.onstr.,a[e abseric:e of dit-lb:;is.an of ihic I.1'':C /DEA:D' assay (i.e. dea7ron;t.ratC 110 tiPODUÃrrCOUs leakage of Ã;herTiic;als}
froiir loa(ie({ narrol.iter coratagne.rs, .ir.a the abscrrce of RF
.rad.iation. 'I"~ie e\~perinrerrt followed t}re test procedures wit1a: the exeeptaora that the RF was raoi iur-raec1 orta. "['Iri.s helped to etisure that the iii-tre frarrae f~'br exposure was the 5arrie.
Confocal rnicro;~~~py was used i:c) verif:y tEa.at: rro le.a.~~~ge, of tE-ie Live/Dead assay E-iad occurred frorxi the 1'~1:(=f'Am in the absence of RF radiation (remote heating) over the tirn~
scale of the exlaerÃraaerat (Fig. 1. 5).
Excrli../c, 5: Sur~ ~~c c> ?'ensiran Dravc-m Se1=FoNing Polvilcr~ra Fabrication ca f'patternect pofj~h~.~dr=a:
1001421 "1`lae t:inst step in the process irrvolvecl t:tac fabrication of 2D
templates c>orr-ilatased of patt.erriec{ faces and aoltier- lai:riges tlaiat woLr:(d eventually fiald iap inÃo 117 ho11o`v polyheclra. A polymeric sacrificial Iaver rr-iade of polyr~-ietlryl meÃIraca-ylate was spin-coai~ed oÃito a silicon (Si) substrate to iaci(atate a{ibseqtierat release of the 21) tezriplate4. A r-actaiiic seed laver was thea7 evaporated orito the sacrificial layer to create wafe;.r-sca:(e electrical contact ftar subsequent e(ectroÃlelaersitioi-i steps. The faces were p,atterrred using phr.rtcalitho~,s.raphy wid fabricated Uti11a1"
e::iect_rodepzgsitio.ta. Since conventional p}rotnlithograpE-iv was used to pattea=aY fiices, any arbitrary patierri could be .ine:orprarai:ed. Faces ccaFnprs:;ecl of eitlrer- copper (Cu) or rilcktl (M) were f_abricated;
c1:roice of n:retais was determined b~,,r cost, otcli selectivlty rn<ith respect to the sce-d lbiyer, ease of depositi t.a5 and the need fÃsr magnetic functionality. A 5eccrzrd ltaye.r of pIotolftkaosMrqplay v:vas tised tt) patiem the solder }ri~ige t. rlapl3tes.
After laingge patÃerrairag>
the exposed seed layer in the la.irzge re~lori kac3Ligaded by tEa.c faces was etcliec:i to cliscontlect tlac wrtieriy'.irrÃ~ seed layer oxrly between tl-ac faces, wfiile retainingr clectrical corrtit-wity Wit}r the rest of the seed laver at tlae face cc?rners. The solder hinges weru electr~~~eposit.ed, and then the 2IJ terr~jfla.tw was rc1ea`es:d I`roi-tr the substra:te by etcliirqg the reraiainirrg, seed lz7yfer-at3d ciissolvint, tlac sa.crifrcitrl Iaves. A tt,raapl:rte conrpt?sGt1: of six sqaarv faces, arrat-r~U~. in a c:raiei.ftrr-rra and held together by solder lairr~~~; ,va5 tisGt1: to form a crit)e Apart frorn the solder iaA between faces, there is no other tether. Self-folding was carried out .in a h.igh bc]ili_iÃ~,,, lac]iz3t so[vei-at, N rneÃlaylpyrsoi.idt>iae (i1;M1?), wlaicl3 tuas heated abave the iTicltirÃet poil-a. of the solder (-188"C) A small a_t-noua-1t of 44-5 RMA (rr)sin, rnfldlSf activated) f-ltix aA-us added to tlie solvi.nf to cleati and dissolve any oxide layers c~i-i t:hesoldvr and Ã.IiEreby erIsÃIre 4~c~c~~~ solder reflow.
13e:sigii t_onsiderati0ns:
1001431 lii tlic design, Ni was a1wavti used as the topmost surface layer of the face in contact wit}i tlie }iiiiges, Even for the Cta poIvliedra, the top ofthe faces were coated with a tltin laver of Ni prior to hinge depcAsitiÃ}aa, Solder does not wet Ni scÃrfa~~s w-ell, so the solder stays in the region where it is elc ctrodeposited. ancl does not spread across the eiitire surface of the face during ~~ldit ~~ (wliie[i occlirs wlien solder is in contact with Cti). When ttie Ni coa:tiiig was absent, we still observed folding, however the yields ,,vÃ:re pcscgr. The low yield was a resiift of the solder migrating away ~rcim the regions vvhe>re it was cl.cpositccf, thereby iiaaki~ig it very difficult to control. the vol~ime of solder in the }iinge region between faces (which ultimately determines tlle fizial ~~ldi~~g angle).
1001441 The design of the 2D template ir.Itimafely determines, the final shape and porosity of the polyhedra. S1iowti in Fig. IA is a typical -2D layoÃit of the tarcs ai1d hinges. AÃÃtodcsh AtttcsCAD 2005 was iÃ.SeÃ.l to generate the lavotit file L~~ed to fat?r:ie;aÃe two pltot.omtÃsk~ (one for the fzÃces, otte for the hinges). To fabricate a cÃÃbc, scfuar:e.fiÃc:e:~ separated b;~ ~gap, c, of 10--1.5% of the face d.in1ension, I.:, in Fig. IA, were t}'t.~ie;alk= U:~etl. SÃsrne tolerance in the gap width was observed, as the molten solder teiicls to draw t(ic fac e;~ laterally towards ea c;h other clLiring foldill.g. l:t shoitld also be noted that sir.aÃ:e the ~.~a}a wiÃIt:(i. .is I0--15% cxf :[.,, it was oftei-i tlle fniD.iITiÃ11Ti feature size of the pbotonnask and Iithcagi-apliv pioc~.`sk, e.g. i-~.~r .15 Lti-ii cubes, tl-ie requirred gap 3vidth of 1.5-2 ~um represented the st-nallest lithographically patterned feature.
1001451 :1n contrast with prior swrfzÃce tension-based seli=folÃ.ling, ovorlti, two types cal 1ii-Yges tVCT'e ÃISetl: internal ones betd~~eeii faces (folding hinges~ and external Ã.anes at the periphery of the t.'aÃ;es (locking biatges). Th.ae folding hinge witll}t (s:liÃ:AN'tt as W irt Fig.
11:3) was '?5% of L, artd the ht.ittw;e 1e2~~Ãh was 80r90`?i3 of I. If tE-ie folding htirlge lertgtlts were smaller the cÃrbes Ccst-a7ed were, iaot seal~d cotitplet.ely at the cÃarrters.
Longer htirtge lengths (>90%) were ut-iÃiecessary, sirtee rleigE-lborirlg hinges would overlap at t1ie corners. AdditiotiaIly, hinge letigt}ts of 100% were irto(artipatible wit}i the tabrica:tiora proÃ:.ess: tliese liirtge patterns Ã-esa.iit.ed in the Ã:,ot-rtp(ete removaI of the seed IaiJer at t:Ei~ peritiieter of tEic 2D templates during the etcb stop after Phtotnliih7o;.~rapE-ly of tl-ic laees. "1'his .retYtoy:t.l -forÃaYerl in art eleetrically discorttitl~torts ..,eed layer that prevented sul-iseqLient electrodeposition of t.hte hinges.
Re~o-w. of the folding ltirtges providecl the torque to rotate a~~acerrt faces. Lockxrtg hirtges that I-tad the same letigtli bLat half t_E-ie width of the to1ditig hinges played a secondary role in tlte foldino.~ (yl: the ~:I) tetr~~~latei thev as a stabilizing stop, increased fiartlt tolerance in folding, atid ertsa.Ãred a 1'atiat fold angle of 9V [ Sytits, R.
R. A. J.
~~,fic~rmd -evoorrtech. t. 1995, 4, 177-I841. Additionally, IÃsckirig hinges increased the t3=tcchtanictri strezt4xt~ht and se.ttli.d Ãhte edges of tf~.c t~st~f~>13~;~clr~a when ~-~~~o htali=<:ire~i l~~clÃ.ixt~;
11in~~es tÃtsed arid formed a. sin<~=te h~i~~e cotitait~ing the equivalent volume of a folciinQ
hinge. t=otdiÃtg was Ã:ott:tplete withzti-t seconds vdtest the 1oi<kistg lti''. ~,=es iitei kintl itFse~~ ~~~ttr each other. The fusiort oecut=ged as a resfalt of Ã1le tYtitiitYtizatiÃ.>tr of interfacial free energy beto~~eett the i-nolteÃt. locking solder bitt:;e on each face 3t-i~.-~ the surrounding lielrtid. 0-zt coolitig, the solder hinges solidified aiid the polyhedral st.ritc.tt.tre s~-as locked in to place.
Finite cleraietrt sintu tal.iotr t;
1001461 T:n orcior to better understand the self=assimbly, process, we perfo:r~~ed titiite.
e1enient sirt-tul.atiun5 crsig-tg the software prc3gr~n-i SLtrfac:e Evolver [Surface Evolver was developed by Kera Brakke -fi`oart tl-ie Susquehanna University Del:tartmertt cyf M,ithenra.tics. The latest Windows version v^.`26Ã;, Ãtpdat~.~ci.
Sehlteiitl:rtvr= .I T, 2005 was ttsr d]. Suafa~~ Evolver determines the tt-iirtimttm energy sutfacÃ; for a giveti. initial sLtr:face atid a set cyC physical ~~iistraints, suc4i as gravity, detisit.y, artcl sÃÃt~a0e tension.
The iterations for evolviazg a mininaÃa.na sÃga=f;tce are cctrttrctlled manually by -thie user.
Scripts were devÃ:lopeti to automate the task of varyin~.F parameters and evolving riireltiple surfaces. Sinau[Ãrtions pe.r.{=ornaed bave :irac:irzcled only f~~io acljjacent. SqrÃare taces, kield to~.~ether by a siriM,le solder fo(d:irawY hing;e, since this cal,tr.Ãr-ed itie essential frÃrietioti of ilie folelir~~.~ tÃinges, that play the ct-iiieal r=ole iri I~brmÃng i well-folded str-uCtUre. (hie face was asSLari-ied to be i^'axed, while the other was allowed to rotate frrvely around the solder hinge; tli:iS assumption parallels what was obServed in eNlaeriÃiieÃits. 'I"~o dete-r-mine ttie equilibrii-rm RoId a:ragle for a given geometry, we used the fiiol1owitig strategs- 1.~-i~.r:~t~, K arid Lee, Y. ~ =:
~'~=~~c~c~~ec~~rr~~s t~f Sf'.~1:; Sari Jose, tJSA.
1E398]: rrÃiniÃ~ia1 energy surfaces were g;eriera:ted t`or- arigles of rotatiori (out of the "D
plarie) between 0 (flat) and 120 (overfolded) in itiereriierital steps of 5 Tlle equi(:ibririrn angle coÃÃesponding to the g(obal mxnimrrm erxerg ~' was then tleterr~,iiretl frr~sn7 the rliiriiÃ-iitiÃ~i of the surface energy trend lii-ie versus angle plot, -for- a particular given face dr li3 E:11tiAoI:'l.
1001471 ShowÃi in Fig= 16 (B-F) are illustrations of ttie firiite eleriieiit simulaticyn f(yr the foldirig process. In ilie 2D teÃnp(a:te, the fo(dzrag hinge solder- iS iti the fc~rrYi of a "I'-sIiaped ri~.~E-It prism. On reflow, the solder IicILie#ÃeS arYt~ forrzis a Ã~ou-rided contour (Fi(I.
I60. DtÃc to the }iig}i interEacia1 teiisiora of the liquid solder (,..
481Ãn.1im) [White, D.
W. G. D-cxns. 1971, -~4~}t~7-3t~;~~~]. tliere is a st~'oi)g d~'ivi~~<~ fork:e to nunimize the e>~pased ifÃterl'aeial area bet-v~,teri tlle rYÃolteÃi solder and ttie srirrow3diÃ-r, t~ufdic licittid, 'i'his dririÃ~F.~ force cÃatayey the solder to ball rÃp w.hicb cesuÃlts in the rotation of atjjace at.
faces, 'T'he fold angle is primarily controlled by the solder volume. We observed evidence for this coeitrol M botl-a simtilatioais and experimental observations.
Different solder volumes gerieÃ=ated underfir?(ded (Fi~. 16 D. ( i ), correctly folded (Fig. 16 E. H), or overfolded (Fig. 16 F. 1) structures. A plot of the dependence of the fold ~iggle on solder -vol.ttme (generated by siÃiit.rlatiotZs, Fig. 17) shows that ttie fold angle decreases witls increasing solder vo1urage. ~xper-iÃ-nerltally, ttie solder vo1Ã.rÃiie Ãhat deterriliries the equiliba=irÃÃn fold angle w.as n~anipulated by coxitroIliÃig the lleight of tl-ie electrodeposited solder for a given lainge geometr),.
1001481 Sitice the scaling properties of the prot.e<sti were of interest, the gravitational potential eneqgy of botti the solder and t(ic faces were taken into account, in addition to t}ÃC iilterlac;ia1 IRÃ1-faCC Cr.aes-gtr ol'the soider. it has beeÃà shoo-~,-n by cÃthers and verifie;i-1 in our simulations tl-iat: Ãbe mkgnÃtude of g;rav:itat:ioaY,t.l e1=f=ects are esyeniiadly negligible compared to the iÃitcÃ-facial sLÃT=Cace eneÃaj~tÃtitii si-r.e-s become large (i.e. ttiÃià scale).
However, oa:Ãr inclusion of a o;ravitational ener(,w tet-tzi aIlowecl LÃs tc) determine the relative magniturles of each of the forces as the feature sizes were scaled up or dowÃt.
The fact that stÃrfac.e farces scale favorably witlt decreasiÃig size is aÃi attractive feature of stirtace teiisioÃi drzveii self-assembly aztd has the potential to provide widespread LÃtiliiy Ha tl3e assembly of microfabracated ia-iic,i-o aiad minz~sc:Ã1e siruct.tÃ:res.
(00149] t'Ãt order to determine the effect of size scaling ort the foldiÃlg process, sim.ulations for ':[) templates were performed witl-a taces sized ti`c?Ãxt the iÃ-iz~ scale to i}Ãc .ann scale for a fixed solder vc31ijÃiie. :(n cacb case, all diÃ-iieiisiotis (height, widtli, arid length) were l.ittearl~~ scaled 1~~~ tlÃc same c~~Ã~.st:~.tit i=act~~r. ri.it ~eriet=;~~~ landscape was ol3sen,ed (Fi4,~. 18) which drives the folding process art~l that tltere are different energies for differerzt fold atÃgles (for a ;;ivetà geÃaÃ~~~~rY. aÃ-id solder voluÃite). '1'he iÃiitial slope of the energy cLag-vcs isÃdicat:es the rnag;nÃtude of'tlÃc rotatiolial force of the faces and dctermzjics w17ether the f.'oIdiÃag process is spontaneous or not.. A
Ãtc(;at.iYe iÃlitiai slope (Fig. IS, 50 tim tc) '2 nini curves) restilts in a spz3ntaneous folclirig process while a positive iiiitial slope (Fig. I8, 4 itim to 6 Ãiim ctÃrves) indicates a non-spontaneous prcicess. The minima in the ctÃrves (Fig. 18, 50 tiiti to 4 mrti) around 100 are indicative of a stable, ecltÃilibÃ-iEim folded c.onflg),:uÃ'ation. The absence of a minimurn in the ctÃrvc for 6 mm, faces implies the absence of any stable folded configuration, i.e.
the two faces prefer to rema:in flat. 'I'}icse results c.ait be explained by noting that as the :~ize of the faces increases the weig}tt increases and gravitational forces begin to dominate ctÃmpaÃ-ccl to the surface tension forces in tlie mÃ-n size scale.
Hence, the initial slope of the ciicrg ,y landscape bccoiiies positive in t.hc mm rango. and the process becoÃiies noti-spontaÃteoa.is. At sÃxialler sizes, sa.irlace forces overcome gra:vitatianai forces and the process 1ieccgmes spontaneous all the way d~~~~~~~ to the Panosca1e. lt.
should be noted that in the siÃnu(ations, bulk properties for the materials and the solder were asstimed and c-AYUZ.ts such as phase segregation, inrcrn-Ãetallic formation, and.
d.iffiision `vi.tliin the solder ivere i~.inored, if't}iese atiaamption,,;.:
Et.olrl, it appears t}tat the self-folding process weatald work oÃ-a the i-iatioscale. For our stan~ard-eonletry, material Ã~ensities, atid solder surface [ensiOft, our SiRIiy~atatxa~~ ~~~ow tlie n1ax-IT11t1i-11 sponNne:,e-)Us t:c]Id.iaiÃ~ si~e to be L ;~= 1400 ~rrn_ S.iina.iia.taoais tilso slic?w that iii aii extreme case of zi Iow, surface energy hinge (10 Ã~yriesic.an; e.g. a la<lit.id polymer) aiirl tieavv faces ('0 g/c.an, e-.Q.
a dei-ise iaieÃ:a.l), folding is still spontaneous for poiyi-iedca as large 3s. 165 una. This implies that it Sliould be possible to fold structures witli taces cozripcasecl of alniosà ~iiy solid material ancl aYiÃla liii-ages ctsinposed. of vai-trially any liquefiable iYaat:Ã;'ria1 up to a size scale of ai'oL1t'2t'~ 165 pt17 for our particular ge<3J11et.[y.
Experimental results:
1001501 Experimentally, cubic poly[iedra rarigirtg in size from 15 pi7a up to 2 i7am were folded (Fig. 19). We }aave also been able to fi-Ad polyhedra of other sltapes. (Fig.
19C). Although we believe smaller polyhedra c:ait be fabricated, we(tave been limited by oter photolithographic capabilities. Below teiis of microiis, hinge gap width$
approach the sub-micron size :scale and alternative pattemitig techniques, such as electron beaiit lithograplay:are aequired to fabricate the 2D templates. Our theoretical simulations show that folclifl-ig of smaller poI~~~odra are spontaneous dLie to the large nlagtlittÃde of the surface forces at small size scales. Although silnulatioiis show tllat the fo(cling of polvhedra with large faces, i.e. 2 mm taces, is a non-spontaneous process, cxpcriiiae:atfi.kdlv w~. -k-vt;re zil_ple to -folt.l 2 ttam cubes. We rationalize tl-iis result based oii i-wo observations. Firstly, agit~itioii ti-rie to convectic~~i currents iri the lteat.c;d fluid occurs experir~~entally'. This agitation can provide the ini.tia1.
cltiviaa.g force tr.Y lift tae~.'s anaa-ginal.ly over the activation barrier for folding, Secondly, it shoL7ld be noted that `vh.ile ,ve proportionally scaled al1. size a=ari;rbles in the simulatiOD;; (C.g. a 2 MITi face was simulated wit~i a [bickness of 80 Pin}q it was tiot possible to do so e\perhnentally. Due to restrictions on tlie height of the photoresist and resolvable aspect ratios, we, electrodeposited a Ãbit.l:ra~~s of onlv 12 ~im for 2 Tt~iii ctibeti; the faces of the experimental templates thus had Ri substantially lower weight, i.nÃ:rea;:ing the threshold at which foldc~~~ ~eca:tne non-spontaneous to larger sizes.
AccotFrtti~g frai= this fixcd:f:ranYe tlYicl:~iess in otar si.nnuiation, .vc determined that the (str¾~e:,t size.to.r which the sei.f=.fioltli_11~ PFOcCss WOtIld ~vork ti~r tI-ac rnateri.t.is rt5ecl in our process is --:7 zmn_ A1tboug"1-I
we do taOt ea:t7eCt t-O ttse ',t lithrxgrapbic prrxc~ss, to fabricate ;trttcit.tres as larye 7 miii,, the process of seltrfoldin4x may still be refe~~~atit: ttà this ;~i.~~. scale, ~;~s~,~;~cirtJl~? in the i~ackrt~~it~~7 of elec:.t-rortic devices.
Tolerance of the prUtt'ss:
1001511 Wafcr scale patterning of the 2D te~~iplatc:s is highly parallel, O.g.
we t.~i.ck approximately 1000 (f.-l00 ltm) aaid I00,000 (.L='i 5 ~tm) w.D crttcÃftarrtxs (yt) a 3"
water. The fo1dÃng process is also highly parallel, and lat;;e nitmhers ot~:
2D templates can be foltlt;ci at unce. Experimentally, the t'olding process also appears to be considerably fault tolerant and we have otteti been able to aciiieve yields in excess of 90% a:tid t`alaricate large iittmbers ~~f polvhedra (Fig. 20). We lia~~e also observed that tc.aldin9 occurred even when hinge regibtty. was not pertectiy, centered across adjacent faces. Experimcntally, to increase fault tolerance, we ta.rgctecl ottt= solder voitime to result in a slighà overi'o1d (.._.100' of rotation trotn the hcarizortta(}.
Since we tt:sed.
locking ltiti<.~es, Ãl~is c~vert~~id etis~tred that the faces tiiet, allowing the locki~.~; hiÃ~,9es to ftrst.'. This increased the tolerance of the process [Svriis, RRA, J.
NIrc~roelc>c.rtrc~inec.=h.
1995, 4, 177-184] and sealed the cubes at the edges attd comers. Additionally, convection currents existed in the tiot scaltzticati clttriti{; the foldisig process.
J'hese c.otivection c<t.trresits wtt;witztt4'cf the 2D tc;ni~~latkas and. Ltic:rt;ases:i fo(ding, angle tolerance by encou.ra~.-ing the edges of tht:. fiact:.s tc) coll.idc., this allowed the locking soldei- liinges to fttst' and liolcf the faces to4~ether w=ith c,ottsiclerable strert4=th. CJacobs, H.O. et al., Science 2002. 296. 323-325-~
Gractas, I3.H. et a(., Sf-zr'nf'c' 2000, 289, 11 70=1 17?1.
C[1nCluSi {)Td S:
l001521 Irt conclusion, a surface tcrzsiota based fe~ldirtd; process has l~aectt presented that can be utilized to t`abticate utii:ct:E-ie.red, hollow patterned polytiedr-a wit:li a wide range of sizes from t:lie mm icp the nm. By leveraging wcl(-t;wtablÃsht;d lithograpbic methods in microelectronics, this faEaric.ation process provides ',i rOtrt.C
tO irI~:.Orptsrale precisely ~~igira.ecred monodisperse porosity, transistors, sensors, and otl-rer information processing devices ori the prxlylicdÃ-a to create "st-nart particles." ITsirig sira-11.ilati011s, we N~ave also demorasÃ.r=aicd iliat the Neling WOr.aid work e.vitla a wide range of t=ace mate:r.iak arid liquctial?[e, hin0es. also dernorastrat~s that the Lrti:liaatiora of inÃed'acial forces, which scale fa.vcaaably at small sizes, is a useful paradigra. for raiicro arid raaraatabri cat:i on.
E a.atn Ve 6: Spatialty Controlled Ch~.~tnist,~~~ ~ ~ing Renaotcty Galhl~.~c1 A'anof.zter GS'c=ale G~~ituhierv 100 1531 Along wit.la coraverational channel based rnic.rntllridic devices, sev'eral nanoliter scale claemical encapsulants have been developed, includin~. those based f~rl pc?lvriiers; gels, and litlr.rid drops [for example: (a) Lim, F.; Sun, A.. M.
Science. 1980, 210, 908a910. (b) Ch~ng, T. M. S. Nin Rev. Drarg Disc.ovcrv. 100", 4, '1l-235, (c) Langer, R. .3.cc. Clarrm, Rcs. 1993, 26, 537-42. (dyfice, J. D.; Sorig, H.;
Lyon, A. D.;
Ismagilov, R. F. Laai(ir~7uir 2003, 19, 9I27-9133.(e) Harnmer, D A.:
DiScla.er.D. E..
Ann. R.c,z=, I.Mati.r. Res, 2001, _3I, _187- 104. In cot rt:ra.st to the above organic systems, nzicromachinecl si1icori-based devices can have extreme prccisiori, high rcprodtrc:ibili.tv, excellent mechanical strength, good chemical stability, as well as the ability to incorporate sens.ira`s, sik;nal c.ond.itionfla;F, and actuating ttanct.iOD;; iD clOSC
prc.e:si.rni#-y oi- ori the saane substrate. H:txz~evcr, :;1:3 i-nic-rotnac1}iiacd nanoliter scale reservoir systems 3vith c:oritr-ollc (l porosity do not exist at the present time dLre to the inherent t.wta diriierasionality of t:(-rc photolithographic process tlaat is r,rsecl in conventional `ilicoia based r.a-lic..r-~rDachirling.
1001541 De;raaonSt.F-ated here. is the deveIopii-aent of 3D containers witla.
precisely engineered sLer-l'ac:c porosi~, and their utility in chemical encapsulation, gLricl~.~ci delivery, and spatially controlled c.hcmis[r~>. Brretly, the process irivr,>lved t1w photolithographic fabrication of.' a 2D metallic template with solder hia~ges (Fig. 22a).
The 2D template self-assembled into the 31) hollow polyhedron when it wa;
heated above the ri-ic;lti~~g point of tlic solder binges, whe.reiii t.EIC
stlrftÃ.c.C tc.DSiOTi zgf tllC tsl solder provided tl-ic force to d.r.ive Selt-asscn-ibly [(a) Syala, R.R:t.
Yea.[mall. E,M.;
Brig,lit., V.M , Whitesides., G.M. I 'NIEVIS 200-3., 12, 387 --- 417. (b) 1-lui, E. ;1; ; I-~ower R. =I ;Rocigers, M. R.; Uth :lnt. Conf MEMS, 2000õ 6021r607.{c} Gimi, B;
=1_eorig; rF; Gu, Z,;'r'ang, M.; Aa-teriiov, D.; 1;31~~ijwalia. Z.; Gracias, D_ H. Biomed.
Micr-odevice"005, 7, -341-3]. Containers 1~ave been tabrica:t.e~ I.N.1itla clit't'ereÃit shapes aiici volurnes ran~irzo t`rc~iia 230 picoliters to 8 tia~iat~likers (Fig . 22 a-d), I'he fabricaÃ:icat~
process was also 1-iigblv pai-a11el; containers of different sba~~~~ ~~ld Sires coulcl be fabricated in a single process run (i.e, from a single wafer, :1~'~~ 211 e-g).
When the process z~,as optimized yields ranged from fi0-90 o (yielcls varied fo~r difTerent sI-~aped caniaine.r~ depending on tE-i~ ~~~~~~~~er of fb(d:ing f=aces and ttie syrnmei.rN) for a 3-wa:ler, I":1le tii.~~or v.icld li.anite.rs itt the t~,br-cai:iort proc~~~
~~~ere t:lic phoioIit1iog-raphic fidelity in the reg:isÃav ot' hinge5 with respect t.o the tices, and t:l~e volu.me of solder if) t1ic hziiges ~Syms :R. R. A. J. MENrtS 1999, 8. 448-455. (15) De.ri.,. 'I'.:
Wliitesides. G.
M.; Rad}iakrishiiati, M,; Zabcyw. Ãi.; PreÃitiss. NI1'':.App1. Pllys_ Lett.
2001, 78, I7r5r 1777]. Since pIaotolltbcgCFraphic mic:rc}fabaYcatrc}n is highly precise, it was a.lso possible to pafterii one or more faces of the containers with monodisperse pores (Fig.
?22 h-k).
'1-1ie size. of tlie pores fomied was limited by the I.~1i~.~tomaskS Lised (wIlicli in this case had a resolution of 3 mici-otiS). Bv controlling the porosity it was possible to ent;Ãneer the rcaigeni release profiles as shown in Fig, 21.
001 '55,55~ fi'(ie c:onta.inerw were loaded ~isi~ig stereotactic Microilijecti~.~n with a solution cif a.:a;el (or po1~~r~~ea ) at~cl the chemical to be released. When the solvent evaporated, the gel r~inaÃnecl within tlie containers. The cheniieals were released by i.nuncrsing the loaded containers in a. sOlLItiOn that softened or cliswolved the gel (or ~.~~.~lvizicr). Siizce gds 4aiid polymers) are available wit:l~ a wide range of aolubÃIiq= aiid s~
c~lenfng temperatures, it was, possibte to manipulate the clieiiiica.l release rates tiSing different ~~~lventS an~ temperatures. The images shown i.n the paper were obtained tising containers, loaded with a block copolymer bydrogc.l (Pluronicnx-).
Release experinieiits were cl.oa-ita in a. water-alcohol 1.~asecl mc;di~IM (Details in the Sul]plernc.nitary SecÃ.ioazl..By varyiaig the relative poros.ity oia diffcrcntfaces of the container it was possible to get both isotropic. (Fig. 23:a.) as well as ataisotrol3ic. (Fi(T.
?3b) chernical release profiles. Since tlae f'abric.ation process was compatible with a variety of aterials, it was possible to l;al?r:icate nickel based containers that could be remotely guic1ed using Ãaaagtietic fields. A spatialfv controlled. (the letter G --any arbitra:ry trajectory is posslble) claciaa.icaJ reaction was demonstrated by directly releasing (~%.Titing) a p:H ÃzadÃcatoa- phezaolphÃlaalcin in a microwcll i~illcd witla an a1k<alinesolutinn (Fig, 23c). Direct writing was possible by iaaanipulating tlae plaezaolplathaleiat-pluronic loaded coiatairter usiaxga magnetic stylus that uasmt~~~ed tinder the micrc?well. It slaould be nÃfted: that while guided manipulation was clone usita.~.~ a laer~~iatietat niagnet, it is possible to use otlicr well-developed microcoil based raaag.aaei:lc ixaanipulation circ.uits, [Deng, T.; Whitesides, G. N1;
Racllaakriy tnait. M.;
Zabow, C~.; Prentiss, M.Appl_ Phys. Lett, 200 1, 78;. 1775- :I777]', to reproducibly control t:(~c moveraaeaat oi"Ãlae corztaiiiei-s and li~iic,e tEic spatial release o~t.'tlae chemicals witli arl}atrard. p;a.tte ta.s.
1.001561 Spatially localized chemical rcactioias were also demonstrated between mu.ltiplc nanoliter scale containers (Fig. 24 a-c). 4k"hen two c~.~titainers loaded Nvit1i copper sulfate and patassiLim hydroxide respect.ivelv were brou:;lat close to each otEier in an aqueoLis medium, a c1a.eataica1 reaction (to ~orixa copper laydroxÃde}
occurred only along the cciitra( line between t(ic two diffiasing rates; tlic reaction occurred nearer the containers witb the slower diffttsing chemical (Fig. 24d-f). These ex1,eriine;ntz farth.cr demonstrate that the spatial coaatrol over chemical reactions can be extended to more c.ornplek rexaction frotit~ involving mt.tltiple containers.
10015,71 -1n conclusion, as opposed to all organic cncapsia(ants, the comaÃners allow taizpreccclciated spatial coiatrol over tlac release of cheiiaical reagents by iTi rttic of their versatility in slaapes and sizes; aitisotropic l`a.ces; monodisperse porosity, and their ability to be gmdded in micrcgl'ltÃidic channels Lasliag magnetic fields.
Additionally, the nieta11ic containers interact witb remotc electromagnetic fields that allow thiaii. to bc easily d~.~ttactecl tza-id tracked (U5ing magneric resonance imaging, MRI).
ThLis, tlle containers proa=ide an attractive laIa1-Cora7a for e:ngi.tac.e:.rirrg rei-iiÃ.stc::ty guided, spatially controlled clae.mica.l reactions in rYiicrc3t lLiidÃ'c. systems.
Fabrication of tlac MicrOcOnta'sncrs:
1001581 .t1. 5.5 pznwthick Sac.riticia.i 1aver ot poly (meÃlaw 1.iaaettlacrw lza.te) (P'41:M,'L, MW: 996K) [Sigriia-Adlr-icli was ypuii oai a silic.on wa:fer.
On top of -lre PMMA-c.c~~ted waf:ei`, a 15.nna ad}resior3-prÃ.arnÃ:at.ir3g c}rr`crrTiiUM (Cr) l~IVer`
and a 100 rarii conductive seed copper (Cu) layer were evap~.~rci.ted. After tl-ae tlairl fill-a}
deposition, we spin coated a layer of Shipley 41'R.220 7.0 photoresist [;Rohiai and Haa.S, www.ro}uYahaas.ccam J, The tkrickriesS of the photoresist vva5 c.otatrtal:lec1 via the spin speed aÃid bv varyina, tlie number of c.oatiÃigs applied_ :1fter- a soft bake;
the resi,,;t lvas exposed to [iV light using an t:ltra pline Series Quintel ma.sk alip,.raer [QuiÃitel C't?rp., v,ww.quÃntelcorp.c.ona ] arad patterned using a ti`a.raslaarenc,v xnask. After developing the photnresisi; electroclepÃ3sitÃr~~i was used to ~.~row tlre rtaetallic t`ratnes of the znic.rocontainer-s with.ira the plaotc3resist mold to a heiglYt of 6w15 urta tdelsendirxgon t:lae characterÃstics reqa.Ãired by ~~arioÃ.~s applications). We used ccammercial electrolytic soIutions that contained the rr-aetal iÃ~iis of choice ['1"echaic, [ne., www:tec.bnie..cÃ-arar j tca e:lectrodelaoait dikTereÃat Ã-iietals. For the construction of non-mag aaet:ic coratairaer-s. Cu was electroplated, and for ma,xietic containers, :~:i was uSed_ A second round of photolitho.graphy was perforÃaaed in order to pattern the laiÃiges. A laver of SI'R'??U was sptira Ã.~ia top of the stibstrate aiad exposed tÃ.~ tlre hinge mask. WÃdeÃ-, irateà rral hinges were located betweet~ aqja.cent faces, whereas the t}ainrier, external liiriges resided at the otiter edgeS of the frames. Alignment muks were tised to enst7xe aligiiment of the hinges to tlac franie>s of the -2D pre:cLirSors. After tlae. Iringe patteraa;:
were developed in 451 Developer, the exposed C:Li (seed) a:Ãad Cr (adhesion) regions Ãra between tlle electrodelaosited frames were etc}ied using commercial etchants Ã:AP'~-100 for CLI and CRE-473 for Cr jTechnic. Inc., www.tec;hnic.comj). Tiai:'lead (60:'40, m.p. -Y
18~? T) solder [Tc:c:hnic:.q 1:11c,., www.tc:c;linic.c;om] was then electroplated iTitc:s the hinge regions.
The hei~.~lit of the hinges was apprc3xiÃiiately 16 ;~~i-ii. xkt`Ãer the solder electrocleposiÃ:iorl, the photoresist layers were stripped off with acetotae, the remaining Cu seed itid Cr adhesion laverc were elclwd atid the 2Drcp.reeursor teiTipiaÃe composed of rnetal frames connected with qolder hinu;~s was ai-nriiersed in NwMet~iv:1 l;'vrrolidone (N\11)} CSi~,~niar Adlrich, www.sigma-alcirich.coarij to dissolve the saci-ificial 1''NNIltIA
layer a:tid release the precursors frc~~ii the wa:fieÃ~, A1iproxiÃ~iaield 50 precursors izl NM:P
wer~ spread across a s~~iall crystallizaÃion disl-a aÃid a siYiaJl aaYioÃint of 45,' RM_A
flux [1Ã1diiim C'cirporaÃion, www.indium_com] was added to dissolve any solder oxides that may have.foa-med. The disli wa.s heated to 100 'C:` for 3 miiiut.es and then r~:~n-peci zip to 250 <,C' loÃ~ approximately 9Ã3 secoÃ-ids uÃit71 the solder becai-iYe ti-iolteÃi Durin4~: re{'lc?~v; i~- the sol~er .:~~et t~ie top 1aver of metal ofii the 2D precursor, the fzabnoation yields were Iloor.
Solder wet copper well but did tioà we Ni well, }ience for contaiÃxer,"Ii ith C'i.l frames it wa.s necessary to add a tliiii Ni laver to iriiprc~~~e yieids, WheÃi ;s6lder reffinved, [tie niolten solder at the Iii~iges asid generated the toÃ-qÃae to fold the 2l:3 precursors into 3.13 micrcgccgntainers. Upon cooling, the solder solidified anct permanently licld the container ~rwnea togethcr.
Ct~tititinei- Loading:
1001591 Two metbÃsds were tised tc? load reagents into the contaitiers, depending o-n the we;ttabili.ty of the c;hÃ;ia-iicaI reagent ori the c:orataines-. Wtieri the ctic:Ãi-iia;FÃ1. wet the c~iitairier weltq several boxc.s ivc:.re simultaneously loaded by im~~iersing t?ien-i in a drop ol='the chen:Ãie;al i-cagent. The solvent was removed by evaporatioii. This lc:ft behind the Pc?lviller [P( T011iocE~' F68, BASF, www.basif.t.oml soaked with the chemical reagent.
10016Ã31 The 5eÃ:or.ad nic:thod utilized t,,vt~ three-a~~s Newport micromanipulators [Models 460A & M462, uwu.neupor-t.c;otn ] to iiidepei}dei}t1y i:caritrol tlxc position oi' the microc:.untainc:r and the s'=rÃ~<~~~ [World Precision Insrr~r:~~~;r~ts, Inc:. :"~~~,~afi1T~~
Syr:irlgc;, www.wpnnt:.z.ora-ij. The syringe was ou[-fitted witli a 36-gauge ~-icedle [WPI;l 36 f_i-augc. Needle. wwwwp.ii.nc.com] t.o facilitate Io,Ãci.i~ig af the mic:.rOc:.ODtai~-lerS.
Cheniacal Release & ~eaclioaa Specatacs:
10016:11 lted I)ye (Fig. ~~a-b}: Containers tvere loaded witlr a mixture cnriipnsed of 1.6 rnl, i0.26I wYl F DX:{_` Red 40 [:~~~cCo-rri-iieA
& Co., lÃic., www.mccorxnick _corn] arrel an aqueous po(vtireric solÃrt-iorr Ã:.orriposed of 1.0 c, of P1a.rrozr.1c:F68 dissolved in 10 niL of water (18A ?~10) . A 2:1:2 (by volr.rrne) mixture of g1ycernl:ethanol:wate:r was used as ttie diffusion metl:itrrir arrd this medium was added to a small cliamber ccrÃrt.~~iiirig ttie loaded miÃ:.Ã-oÃ:.otxta.itier. The diffusion profiles were irr-rkged usizig a. stereozoom binocular rgzia;roscope.
1Utll.~"i21 caliyr-G uided 1'berro(pbt&ialei.ti-KOt=:l Reaction (Fig. 23c): 'f .lre indicator mixture for the pla~iiolplat(ialein-k.OH reaction was pr-epa:red by adding 0.25 mL of p}rertotp}rthaleirr solution {0.5 h; of ptr.errc?lpht.lraleitt [Frey Scientitie.
~~wv.v .:i=a~evscieÃrtitic .coÃ-ir] .i11 Ioorr-ii. ot ~~5%etlrarloi)toarr afli,ÃeoLispolyri-ieÃ-icsÃ~lÃtt:i(ari composed of :I .O g of PlÃrronic F`68 d:issc)lved in 10 ÃTiL -water= and loaded irrto a Ãrickel-based microcontairrea, The microc~~nÃairr~r was placed irrto a well of a tissue ctiItuÃ-e pIate, [;Falcozi(P)1~.1u:ltiwell`l M`l'issa:~e Culture lslate: 24 Well, arici a 1:1:1 (by vo[Ãr.me) g;lycer-ol- water: I ?~~~ KOtl(aq} mediurrl was iriirodÃtcecl irito the chanil?er. The micrcgccgrit.ainer was guided a-nd, controlled usMrw a 0.35 pcÃli lb., :1 1/8"
dia~icter A_(\iCo rouaid bar magiiet [McMastea -C"arr, ww-w-.mc:ma.ster.coÃr Ãj.
C x~~>~?ei ~ Il) St:al~`i'tt~> ~'c fr~<t~?t'~~t~ate fsl..)t~{ Rccic;ifon ff-ig 24a-ck L'uSC.)::~aq3 + 2 KC.)H(aq) K,S(.).,(aq) + C'Ãi(OH)>4s) 1Ã)Ftl.1.~"i31 The copper sulfate reactaÃit mixture was pr~epar-ed by dissolvirio 1.0 g of Plu.rorriÃ, F68 irrÃ:o 10 rril. crt' 0_5 "~=1==; Cu{ii}~~~~ aqueous yolÃrt:ior [Sit'rrra-Altl,riclr., z~ ~~,w.si_Qrr-ia-a1tir.ic.}i-com I arrd was loaded irrto a ri-iierÃsc:4sit.tairrer. The potassi111,11 hydroxide reactant tiaix[us-e was larelaai-ed by dissolving :1.0 g of P1uronit:. F68 fl-Itts 10 rrrl:, of 1.0 M KOH(~~) arrd was loaded into a second rnicroconta.irrer.
'f'1're micr-ocoÃ-ttairae~~ were placed in close proximity into a poly (dirxretlivl siloxane) [PDMS> Dow C:`orrr:iÃrg Sylgard(k# 184, www_dÃ~N.veorning.con) j trrier-owelt.
'f .lr.e znicro-~x-e:ll was fabricated by:mc~lding PDM:S against aii SU-8 p}rotoresist Ãn. aster: The ditfus'Mr and reaction Ãriedium was water.
1~=f3I1----Rcxcraicm (j_{g '4_ `.l'he iTtc{ic:a,tor ri-iixture for the p1terinlp}rthaleinwkOH i-cactioti was prepared by adding 0,25 mL of phenolphthalein solution to an ac1UeOLIs pÃ~IyrTtcrie sO(utioti eÃ7i-a7pÃ5s-cd of 1.0 g of Plurortic F68 dissolved in 10 txzl, water. The alkalirle ri-iixtu.re was prepared by acldirtg 0_3 mL ol 4M :Kt7H(aq) [Sigma-Alclrich. W-w,~x.sagtnaraldrÃch.comj to an aqueoLts polymeric soltÃtion composed of l,O g Pluronic F68 and 1.0 iTIL water. Two containers were loaded wit:ti the pli~iin11~lit:EialciÃi sc~lutioia azid one vvith Ãbe KOt:-:( solt-itiÃiti. 'The tkirec containers were tlw.ti placed into a 1'D:MS Ãiiici-owe(l, with water as itie diffusion atid reaction medium. 'I'}ie reactions were also imaged uSino a Stet-eozoom binocular micrc?scope.
1[){1164] W1ii1e the presetit: itivetitioti E-iaS bec~i described with re{eretice to the specitie evabod.itx~ents t}icreof it should be understood by those skilled in the art that various changes may be made atid eqttivaleiits may be substitÃtted without departing troti-z the true spirit arzd scope of the iriveritioÃi. :In additinri< many modificatiot~s may be zrtade to adopt a particular sittiat:ioti; ma.t:cr-ia(, compoyitiott of matter, process, process step or steps, to the objective slairit and scope of the present invention. Ali st.tc:li modifications are intended to be witliii t the scope of the c.laims, ap.1,e~~~ieci hcreto.
Claims (52)
1. A three-dimensional particle comprising a plurality of two-dimensional faces capable of self-folding to form a hollow interior, wherein a size of the particle is microscale or nanoscale.
1a. A particle of claim 1, wherein the size of the particle ranges from 1 nm to 2 mm.
1a. A particle of claim 1, wherein the size of the particle ranges from 1 nm to 2 mm.
2. The particle of claim 1, wherein the particle further comprises at least one hinge.
3. The particle of claim 2, wherein the hinge comprises any liquifiable material.
4. The particle of claim 2, wherein the hinge is selected from the group consisting of a polymer, a gel, glass or a metal.
5. The particle of claim 1, wherein the particle has surfaces forming a polyhedral shape.
6. The particle of claim 5, wherein the shape is a cube.
7. The particle according to claim 1, wherein the two-dimensional faces are patterned with perforations or pores.
8. The particle according to claim 7, wherein the perforations or pores are created photolithographically, electrolytically, or by using electron beam lithography.
9. The particle according to claim 7, wherein the perforations or pores have a size from about 0.1 nm to about 1 cm.
10. The particle of claim 9, wherein the perforations or pores have a size from about 10 nm to about 1 cm.
11. The particle according to claim 1, wherein the particle is fabricated from at least one material selected from the group consisting of a metal, a polymer, a glass, a semiconductor, an insulator, and combinations thereof.
12. The particle according to claim 1, further comprising active electronic or semiconductor components including transistors, sensors, actuators, light emitting diodes, photodiodes and solar cells.
13. The particle according to claim 11, wherein the metal is copper or nickel.
14. The particle according to claim 1, wherein the particle is a Faraday cage.
15. The particle according to claim 1, wherein the particle is coated with a biocompatible material.
16. The particle according to claim 15, wherein the particle is associated with a biosensor.
17. The particle according to claim 15, wherein the biocompatible material is a metal, a polymer, or a combination thereof.
18. The particle according to claim 1, further comprising at least one substance encapsulated within the particle.
19. The particle according to claim 18, wherein perforations or pores in the two-dimensional faces of the particle allow release of the contents of the particle.
20. The particle of claim 18, wherein the substance is a therapeutic agent.
21. The particle according to claim 20, wherein the therapeutic agent is selected from the group consisting of a cell, a chemical or biological agent, a pharmaceutical agent, a composition, a tissue, a gel, and a polymer.
22. The particle according to claim 1, wherein the particle is administered to a subject and location of the particle in the subject is non-invasively tracked by magnetic resonance imaging (MRI) or CAT scan (CT).
23. The particle according to claim 22, wherein the particle is imaged with negative contrast relative to background or positive contrast relative to background.
24. The particle of claim 20, additionally comprising a radio frequency tag.
25. The particle of claim 24, wherein the substance may be released upon the particle's exposure to a pre-selected frequency.
26. The particle of claim 12, wherein the substance may be released upon the particle's exposure to electromagnetic radiation.
27. The particle of claim 26, wherein the electromagnetic radiation is triggered remotely.
28. The particle of claim 26, wherein the electromagnetic radiation ranges from 1KHz to 1 Peta Hz.
29. The particle of claim 20, wherein the substance may be released upon the particle's exposure to inductive heating.
30. The particle of claim 29, wherein the inductive heating is triggered remotely.
31. A method of fabricating a three-dimensional particle comprising a multitude of two-dimensional faces that form a hollow polyhedral shape and containing a fillable center chamber, the method comprising the steps:
(a) fabricating a multitude of two dimensional faces;
(b) patterning the fabricated two-dimensional faces;
(c) patterning at least one hinge on the patterned two dimensional face to form a hinged edge;
(d) joining a hinged edge of a first patterned two dimensional face to a hinged.
edge of a second patterned two dimensional face to form a hinged joint;
(e) repeating step (d) to form a two dimensional precursor template having hinged joints between adjacent two dimensional faces; and (f) liquefying the hinges of the two-dimensional template using heat to initiate self-folding;
thereby inducing the three-dimensional particle to self-assemble.
(a) fabricating a multitude of two dimensional faces;
(b) patterning the fabricated two-dimensional faces;
(c) patterning at least one hinge on the patterned two dimensional face to form a hinged edge;
(d) joining a hinged edge of a first patterned two dimensional face to a hinged.
edge of a second patterned two dimensional face to form a hinged joint;
(e) repeating step (d) to form a two dimensional precursor template having hinged joints between adjacent two dimensional faces; and (f) liquefying the hinges of the two-dimensional template using heat to initiate self-folding;
thereby inducing the three-dimensional particle to self-assemble.
32. The method according to claim 31, wherein the hinges of step (c) comprise a material that can be liquefied.
33. The method according to claim 32, wherein the material is a solder, a metallic alloy, a polymer or a glass.
34. The method according to claim 31, step (a) further comprising the steps (i) spinning a sacrificial film on a substrate to form a first layer;
(ii) layering a conductive second layer on the first layer; and (iii) patterning the layered substrate by photolithography.
(ii) layering a conductive second layer on the first layer; and (iii) patterning the layered substrate by photolithography.
35. The particle according to claim 31, wherein the particle has a size that is microscale or nanoscale.
36. The particle according to claim 31, wherein in step (b) the two-dimensional faces are patterned with perforations or pores.
37. The particle according to claim 36, wherein the perforations or pores are created photolithographically.
38. The particle according to claim 36, wherein the perforations or pores have a size from about 0.1 nm to about 100 microns.
39. The particle according to claim 31, wherein the particle is a Faraday cage.
40. A method of imaging a particle according to claim 1 that has been implanted into a subject, the method comprising the steps of:
(i) loading the hollow interior of the particle with at least one substance to form a loaded particle;
(ii) administering the loaded particle to the subject, and (iii) noninvasively tracking the particle of step (ii) in the subject by magnetic resonance imaging.
(i) loading the hollow interior of the particle with at least one substance to form a loaded particle;
(ii) administering the loaded particle to the subject, and (iii) noninvasively tracking the particle of step (ii) in the subject by magnetic resonance imaging.
41. The particle according to claim 40 wherein perforations or pores in the two-dimensional faces of the particle allow release of the substance in the hollow interior.
42. The particle according to claim 40, wherein the at least one substance of step (i) is a therapeutic agent.
43. The particle according to claim 42, wherein the therapeutic agent is selected from the group consisting of a cell, a pharmaceutical agent, a composition, a tissue, a gel, and a polymer.
44. A method of treating a condition introducing into an animal in need of treatment at least one particle of claim encapsulating a composition, wherein the composition is released through one or more pores within the particle into the mammal in an amount sufficient to treat the condition.
45. The method of claim 44, wherein the pharmaceutical composition is contained within one or more microbeads.
46. The method of claim 44, wherein the condition is diabetes, and the composition is one or more insulin-secreting cells.
47. The method for imaging a particle of claim 1 that has been introduced into a mammal wherein the method comprises using magnetic resonance imaging.
48. A method for targeting the particle of claim 1 to a cell within a subject comprising the steps of:
a) attaching to the particle an antibody against specific to the cell;
b) introducing the particle into the mammal, wherein the particle is targeted to the cell.
a) attaching to the particle an antibody against specific to the cell;
b) introducing the particle into the mammal, wherein the particle is targeted to the cell.
49. A method of delivering one or more particles of claim 21 to a subject, wherein the particle is programmed to remotely release one or more reagents at a specific time and a specific location.
50. A method of claim 49, wherein the particle is remotely guided and imaged using MRI or CT.
51. A method of claim 50, wherein the particle is capable of releasing a content agent or of providing contrast to allow MRI CT imaging of its contents or of substances within its vicinity.
52. A method of conducting non-invasive biopsy or microsurgery, comprises directing the particles to a site within a subject using remote means, allowing the particle to capture one or more substances from the site, and obtaining the substances from the particle, thereby non-invasively conducting biopsy or microsurgery.
Applications Claiming Priority (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US81606306P | 2006-06-23 | 2006-06-23 | |
| US60/816,063 | 2006-06-23 | ||
| US11/491,829 | 2006-07-24 | ||
| US11/491,829 US8236259B2 (en) | 2005-07-22 | 2006-07-24 | Self-assembled, micropatterned, and radio frequency (RF) shielded biocontainers |
| PCT/US2007/072029 WO2008108862A2 (en) | 2006-06-23 | 2007-06-25 | Self-assembled, micropatterned, and radio frequency (rf) shielded biocontainers and their uses for remote spatially controlled chemical delivery |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2656648A1 true CA2656648A1 (en) | 2008-09-12 |
Family
ID=39738947
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002656648A Abandoned CA2656648A1 (en) | 2006-06-23 | 2007-06-25 | Self-assembled, micropatterned, and radio frequency (rf) shielded biocontainers and their uses for remote spatially controlled chemical delivery |
Country Status (4)
| Country | Link |
|---|---|
| EP (1) | EP2037809A4 (en) |
| JP (1) | JP5451385B2 (en) |
| CA (1) | CA2656648A1 (en) |
| WO (1) | WO2008108862A2 (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2013013328A (en) * | 2009-10-30 | 2013-01-24 | Univ Of Tokyo | Device having foldable structural body |
| US9630178B2 (en) * | 2010-03-17 | 2017-04-25 | Ut-Battelle, Llc | Method for preparing small volume reaction containers |
| CA2883080A1 (en) * | 2011-08-26 | 2013-03-07 | Vecoy Nanomedicines Ltd. | Pathogen and substance traps |
| US9058552B2 (en) | 2011-10-26 | 2015-06-16 | International Business Machines Corporation | RFID tag temperature adaptation |
| US11331085B2 (en) * | 2014-10-16 | 2022-05-17 | The Johns Hopkins University | Bioresorbable self-folding tools for surgery, single cell capture and manipulation |
| JP6786120B2 (en) * | 2016-08-17 | 2020-11-18 | 国立研究開発法人物質・材料研究機構 | Cell encapsulation method and encapsulating cells |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US2369392A (en) * | 1942-07-21 | 1945-02-13 | Nat Folding Box Co | Hinged cover container |
| GB9203037D0 (en) * | 1992-02-11 | 1992-03-25 | Salutar Inc | Contrast agents |
| AU2006272749B2 (en) * | 2005-07-22 | 2011-09-08 | Johns Hopkins University | Self-assembled, micropatterned, and radio frequency (RF) shielded biocontainers |
-
2007
- 2007-06-25 JP JP2009518492A patent/JP5451385B2/en active Active
- 2007-06-25 EP EP07870998A patent/EP2037809A4/en not_active Withdrawn
- 2007-06-25 WO PCT/US2007/072029 patent/WO2008108862A2/en active Application Filing
- 2007-06-25 CA CA002656648A patent/CA2656648A1/en not_active Abandoned
Also Published As
| Publication number | Publication date |
|---|---|
| EP2037809A4 (en) | 2012-11-21 |
| EP2037809A2 (en) | 2009-03-25 |
| WO2008108862A3 (en) | 2008-11-06 |
| JP2009541490A (en) | 2009-11-26 |
| JP5451385B2 (en) | 2014-03-26 |
| WO2008108862A2 (en) | 2008-09-12 |
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