GB2468701A - Cell cultivation receptacle with extending closure member - Google Patents

Abstract

A cell cultivation receptacle (10) is disclosed comprising a recess layer (11) having a downwardly extending recess (12) defining an opening (13), one or more lateral boundaries (14) that taper inwardly in the downward direction from the opening (13) to intersect the bottom (16) of the recess at a bottom layer (17). The receptacle (10) has a removable closure (18) including a closure member (19) which in a closed position closes off the aperture (13), the closure member including on its underside a downwardly extending displacement member (21) that occupies part of the interior of the recess (12) in the closed position, the displacement member terminating in a lower surface (22) which defines a cell cultivation space in the recess, and the displacement member (21) including a portion that tapers downwardly at a more acute taper angle than the taper angle of the or each said lateral boundary (14).

Description

A CELL CULTVA11QN RECEPTACLE AND RELATED APPARATUSES INs rivention n&ates to a cefl cuWvaton recsptace. The Thvenflon also relates to an appacatus ncftthn such a receotacta, ate for ocKnq EteThon n borb-v ooaatuses.

foc3wr the ttre ws with the desired mixture ot cub enoe th te pte.

be tie 3ter eooor1ence wh t wh ase.3nc cutirs furomtrcft. rnter of the tire pate ack9rn fiuoresenc that bcorne statist e when th each ttre we e rdu id c rirrr wfth rQthred when orventc tre Aih t PO$be ahev urab mcr:osn th eq& rmni rur to o so i exene flc:i ompicati, Furthemre. as p *trdi downwardh, rto th we bobbr p. The gth th pton ty es hm the depth o each we, such that when the ccwer s a ema space exss th La rv tkn asoect ts a rovLded, ae cu vatcu rceaac yar LavLq formed therein a downward ectendinr rece. defininc,. r

The sn c a. peiec -a.bunderv i he rec.etece rr en thet e trr o organk ttere tre cu: hqud ecips deafrebk e deta bek.

-

irUcjar ber.flt th (1 ft r th botcm th res s ncavy curved. euch er arr qen the -*-. -.4-'.

and reflected or scattered hght (as ma result eg. from tiuoresccnaiy m the Convenientl) the closure member and the displacement member are integral Wth one S anomr. Thh permits a simple structure that ma be straightfonwardk' moulded usnr per se Known manuiactunng techniques.

in a practical embodiment of the invention there is provided a plurahty of the recesses formed in the recess' layer.

in such an arrangement preferably there is provided a respective saict displacement member for each recess Such a conflourafion may conyenieriti. be provided in a receptacle in which a common ciosure member has extending downwardly from an underside a plurality of displacm members each iocated so as to penetrate a said recess on moving of the closure member to its closed position.

This arrangement i5 convenient since it permits the simutteneous closing oP of a the recupt:aces of a, test plate in such a way that microwell conditions care simuitaneou&y c:reatel in each ot them in another arrangement, within the scope of the invention, however it may be desirable; to provide a piuraiity of closure members each having one or more thsplacernent members or; extendinc downwardly from an underside.

Preferably the receptacle of the invention includes one or more microchanneis nterconnectinc two or more of the. recesses.

to When as soecrtied herein the recess iayer and the base layer are fnrrned separately from one another, the or each microchanne; desirably may be deflned at the boundary the recess iayer and the baae layer.

Numerous oplions exist for crealing the microchannels. For exarnpie a haif..round or; channei may be formed in each of the recess layer and the base layer so that on asse.mbiy of the layers together the respecfive channels may akin in re isler with one another to define a circular crosssection niicrochannei, in another arrangement a cnanr'tei may be formed fag by miitingi in the surface; of one ot the base end recess layers the other of which Is planar to procitte a flat bounder to the nycrocneinnei Conveniently the receptacle inciuc:es. defined in the closure member an observation region at. whlcn optical signals generated in the rnmroohannei are detectable.

Multiple miorochartneis oan Form a nicrbou?dc circuit, and can be used as rrccro4ofai ic ana/ysfof (pIAS) systems such as described in the ournal Lab onaCh4cf CRc at SooIet of Chemistry. 2007 Cwwwrso.orq/ioch. In such applicauons the recesses are used to load one or more microfluidic drcuits in an automated fashion, Opbortaily the receptacle may nolude one or more elements selected from the i5t m comprisin optical sensors, chemical sensors and flow controi elements located so as to act on or be nlluenced by fluid in a said fluidic microchannei, Such features are panicularlv benetmai when considering uiAS system ap pcations of the receptacle of the nventon.

aC Reoarcliess of whether the rmcrochannels are present or not. and reoartiiess of the rnturochanne constructton, when a plurality or the recesses ts proy the prererably are arranged in a regular pattern In the recess layer.

More particularty, the plurality of' recesses may advantaqeousv be arranged in mutually orthooonal rows and columns so as to define a grirHike array. Such an arrangement has beer? showrt to be highly suitable for expenniental purposes since each recess may be identified by coordinates referrino to its column and row whereby assay and screening results may readli be corn ared, stored and transmitted.

ln another arrangement according to the invention, a well may be oroyided underlyno the or a said recess and register therewith. Such a well may optionally be defined by a well laer thar underlies the recess layer and has formed therein the well The provision of a well as aforesaid may he advantageous when it is required to provide an enlarged celi culture space.

Convenientiy the we Layer is transparent. The weL ayer may addibcnafly include }Orrnecl or defined therein one or more optional oplicai prlems. mirrors and/or other opbc elerryants : Such features aflow for alec viewno of Uie wefl. or socaed "dat* fe/cf microscopy.

When piura recesses are provided, preferacly the receptacle nciudes a respechve sale w underlymp each recess, m In ar. embodftnertt of the inventon the thspiacement member includes tormed therein one or more riule flow passa as that permit the conveyance of fluid to and/or from the interior of the recess.

Preferably the or each said Passape terminates in a further openino in the saE iower u5 surface, Convanenuy the receptacle includes a plurality of the passages and a plurality of the furtner openinos, that are arranoed in a reqular or irregular pattern n the lower surface.

cc Preft.rabiy the or each Passaoe is connectable to a source and/or a drain of fluid.

le one preferred arrangement the receptacle includes a plurality of displacement members rom or more of whch each nclude a said Passace and are fiuldicaily connected to one another Optionally the mspiaoement member may substantially Occupy the recess when the closure closes of the opening, in an embodiment of the invention the receptade includes received within the recess one or more spacer members that space the displacement member from the bottom of the recess, the receptacle includes in the recess a fluid of lower density than the matenal of the spacer. This ensures that the spacer remains in piace of the bottom of the recess after b&no positioned therein 1 1 The spc.i member my tnay he kmed from a tr parent matena The ecer or aflemabvev rn ro1ude orned tieen ne or more throuahqon hoies ..

Convr-rt to ths ic reatec ends, tre rec9pL e Opon& cuds o more.

ftd 1ow prt that re fludav con ecthe br su fiui to Iormm the iUd f'ow path. -,

As nc.*: :1 is preferable; that the cannuiae are supported on a common support that s mcveate relative to at least one said recess so as to permit insertion of the cannulae into ano withdrawal of the cannulae from the said recess'. Even more preferably the support su.ooorts a piurahty of injectors arranged in a pattern oorresponmnq to the S pattern of a iuraiiy of the recesses, When the receptacle ndudes a flud flow path as defined hereinabove optionally one or more saic cannuiae n use is tludicafly connectable to the flud flow path, Such an an'anpmen s beneficial when performing a parucle. capture experiment as outhned is hereinabove, Accorcino to a' further aspect of the invention a mefhod of treating particles comprises the steps of: hi coatinq the recess bottom of a receptacle or of a receoracle of an tS apoaratu's as deflnec herein, havir its closure member m an open position, with antibodes: firt causing the closure member to adopt its closed posibon thereby definmo the fluid flow path; (IS) causinp a first fluid, containing one or more particles to whicti the said, antiboc:ies are antioenc. In flow in, the fluid flow oath' dv) causing a second, rinsing fluid to flow in the fluid flow' path: and Nh detecting the particles in the recess, Optionally the method may inciurie the further steps of: (vi) causing a third fluid, containing opbcaiiy rietectabie further antibodies, that are antigenic to the particles, to flow in the fluid flow path, and (vS) detecting the particles using one or more optical detection technipues.

The terms °mia'owair and "rn,w'ochannef' as used herein will be understood by the St work'er oi' skill in the art; and furthermore may be interpreted by reference to the prior art documents menf).onec..

he lisung or discussion or an epparentiy pnoNpuoushed document in des speciticsuon should not necessarily be taicen as an acknowied emen.t, that the document s part of the Sf3 state of the art or m common general knowledoe, Vt Tr ois descrpion (> prtrred mbodmrt9 of tr4 VtK1, by y of flOn11fl1Q mPO benq to tb oompanyro rwno) wh.

I vrth Referuno.o the drawings, a receptacle 10 in accordance with the nwnnhon comprises a recess layer 11 having formed tnereTh a recess 12 thaI. extenc:s downwardiy from an orernng I 3 lormed in the upper surface of recess iayer I when the receotacie 10 is n S ts iii.ht onentaton as shown in Figure 1 The recess includes in the embodiment shown a single lateral boundary 1$ that lerminate.s at ts lowermost extren'uty by intersecting the bottom IC of the recess 12.

o The sinole iaterai boundary (wail) 1$ in the embodiment iflustrated arises because the recess rottom 16 arrl the opening 13 are each c.ircuiar or Ovoid in shape. Therefore a single, continuous lateral boundary 1$ clef ines the recess 12 as essenhally frusroconical n shape; in the embodiment shown, Recess bottom 16' is planar, and extends horizontally although as. described herein other arraernents ot the recess bottom are possible.

The diameter of recess bottom If. is less than that of opening 13 such thar rsice;ss 12 tapers inwardiy in the downward direction when the receptacle IC is orientated as shown $C m Ngure 1.

The recess bottom 16 is defined in the embodiment shown by a bottom lever 17 extendin along the length of the receptacle 16 underneath the recess layer II, The receptacle 10 &so includes a removable closure 18 ii) the form of a horizontally extendin. planar closure member 19 having extending downwardly fncm its underside (that m the cmbothment shown is flat) a dmpiaoemenc member 21.

Displacement nneniber 21 is in the Figure 1 arrangement a c!rcular trustocone. the centre axis of which is aiigned with the centre of the circular recess bottom 16.

Since cnspiacement member $1 is a fnistum of a cone, it terminates at its lowermost end in a iower surface 2$ that extends horizontally when the components of the. 16 are. orientated as shown n. Figure 1 The length of displacement member $1 is such that Wnen the closure member I 9 doses or)ervnçl 13 lower suchce 2$ lies a short distance above recess bottom 1.6 so as to define.

a mcrowell cell cuttivation space. between the respective surfaces 1 6 and 22.

As sgnited by the symbol u" in Figure 1, the exterior of displacement member 21 tapers at an anqie to the vertical that is less than the angle i subtended to the: verboal by the; latetw boundary of the recess 12.

As expianed, tds feature ensures firstly that even thouph the displacement member 21 O tapers in a sirollar tashion to lateral boundar 14, a gap exists between disolacement member 21 and lateral bounder's 1$ all around toe interior of receptacle 12.

Various advantage.s of such an arranoement are explained above, IS T'he closure member 19 is essentially planar on its underside such that when placed in position to close off the opening 13 it seals around the periphery ol the latter. This mnimises the risk of evaporation of llquds contamed withn the recess 12.

The seaiin also n'Unimises the nslc of cross contamination between. multinie recesses' As is apparent from study of Fipure I lbs angle subtended by the laterel boundary 1$ relative to. the recess bottom 16 s equal to (9(1 fS) degrees. In the embodiment shown the. subtended angle 90 � fi lies in the range 170 100 degrees, and espec:iallv deprees.

Typically the depth of the recess layer 11 is in the range 100 mcnanietre " I cm, and preferabty is between 500. r roo'srer and 5 mm. The most preferred thickness is in the range 1 mm.

so The fl'nckness. of the bottom layer preferably is 1 nanometer -3 millimeter and preferably s between 100 micrometer and 1 mm A particularly preferred thickness tbr the bottom laser is 160 micrometer.

The components of the receptacle io may be made from eg. glass, quartz, mica or a polymer material such as polypropylene, polycarhonate and cyclo-olelin Polymers, polystyrene, pcinathane. PDMS' and similar matenais. The light transmission r(;tetcs th ri teria mv b to cOtior.

the hwr', mnt member 21 s ftrme searat&y from the t' The ocbcal properties of the receptacle 10 can however be further unoroved by usiop one or more orThonal leatures as shown in Fure 2.

in Figure 2 a receptacle 10' is shown that in many respects is smliar to receotecle 10 of o hgure Thus ri Figure $ the recess layer 11, opersrig 13, closure 1$ and closure member Itt are e.ssentia liv the same as their counterparts in Figure 1. On the other hand the shape of the recess 12'. lateral boundary 14'. recess bottom 16' displacement member 20' and lower surface 22' differ from their counternarts in Figure 1 and therefore are represented n Figure 2 through the-use of pnmed numbers, More parbcuiarly. the lateral boundary 14 is not straiqht-syallej when viewed in vertical crosssection as shown in Fgure 2. and instead is curved.

The curvature cd' lateral boundary 14' in Figure 2 is tur$darnentafly parabolic. althoucih other types of curved surface maybe employed. The use of a parabolic curve however is desirable s-moe ts dimensions are easy to calculate.

o The recess botlom 1$' is curved n a con tinuahon ol the curvature of the lateral boundary shape 14'. Since in preferred embodmients the recess bottom 16' is made of a transoarent matedal this I orrn of recess bottom acts as a lens the optical characteristics of which can be optimised to suit the measurement apparatus in coniunction with which the receptacle 10' isemployed.

in a similar fashion, the displacement member 21' includes a lower surFace 22' that also is ourveo. gerienaily m congruity with the curvature ol the modified recess boltom 1$'.

The iransrjarenc of the displacement member 21' and closure member 19 means that :o the closure 1$ atso acts as a lens.

rhe-curvature of the recess bottom 16' in addibon allows for collection of the particles 23 of interest in the lowermost pan of the recess 1$'.

so This can lead to a more controilec: concentration of the' rarticles 23 in a preferred part of the. recess 1$'. Figure 2 schemahcaliy shows a higher concentration of particles $3 at the wemot part of rece 2' than r the c reqkm of recess 12 o The uratue of owerrnot surfece 22' of Fure 2 euses efrcor of ridert " n-'-' . er 24 ni b. manufactured itgray with either or both of rce cr11 and bottom rri. VVe ayar 24 may be made from e. uaft:.. a &oh Tti r c:roud; r'ne may ha brmed e g. by mUllng, athfr or otherwaa formn rtr aq, samroun rer channes the undarakia of eath ca wer I I ana arface of the botti vr I 7 se < or more do rods & that *nerae rough fte ooe member I 9 and tess ayer I I o n me m ofludo canne. onneoton of periorate the. closure member 19 oF each receptacle 10 for the purpose ofi ecbno limb nb each reoess 12 anwor removino fluid therefrom.

in another arrangement a plurabty of the receptacles 10 may be moveabie tateralfy so as s secuenhafly to underHe an ineotor $2 that is constrained to move onlv':n the vertical direction. This represents an alternative way of achieving multiple reo.eptacje dosing.

injector 4$ is shown, in schamatc, form. although rt may adopt any of. range of practical formas that will occur to the worker of ordinary sKrI in the art. Such formats irickide multioh4iead iniectors. that are capable of inptcting plural numbers of the recesses 12 smultane.ousiy in the arran ement shown in hgure 5 the recesses 12 may be preseaied by way pf the closure members 19 in practice a common closure member would be rirovided for afi IS therecessesl2 Perforafion of the 0105u e* nienber(s' 19 by the cannulae $3. $4 niinimises the possibihty ot evapoiator. hum the recesses 1$. As expounec nareinabove$ the nmteriai O tire cJosure member 19 is xselfrseeJing evaporation wouid not occur even after wthdrawai of $0 the cannufae 43, 44 as a result of the vertical motion of the iniector $2.

The irector $2 may be mechancaliy, eiectricaily and fluidicaily connected to a control and supply arranoement the precise design of which will be within the capability of the Figures 6 to 10 disclose vanous optional features of receptacles, closures and anparatuse..s in accordance with the invention.

in Figure cA there is shown in vertically sectioned view a closure; 16 according to the.

invention, that includes an inuse horizontally extendinu closure member 19.

As speciiIed herein, the invention resides in a closure as defined and/br descobed herein when consderec on its own, ie. separately from the remainder of the components of the receptimie 10. Figure BA iS illustrative of this concept; and also shows further, optional at. features ot the closure. Such features may be present either when the closure is considered on. its own oras part o a receptacle. lb. Fjre $A ment member 21 roudes a cce 46 that of shape of th domvardy extendnc. apaomer member $1'; and ar outer we 47, r " rec& 2 o rcpte IC s cred h9refl A& w be ap ien to the worker of the arL a wide variety of arrangements o the fluid ssees and their irterconnetnn, s oossbie within the scooe of the nventon.

to secure the closure member 1$ aganst leakage. Variousniens. wtthn the knwledqe o toe worker m skth ci the art, may oe eniployeu to achieve such seam. As examples one may conscler the use of ciamrllng members. fasteners and adhesive compounds.

o Among other things, the advanta as of the arrangement shown in Fioure 7 are that the well sirnansions may oe chosen to suit the experiment under consideration. Thus, for exam a, if zebrafish embryos are cuitivatsd, the wells 2$ of an array of receptacles 10 according to the nvenbon may be between 0.8 milllmebes and mililmetres in diameter, and preferably between 1 and I.5 rnilhmetres n diameter For such aripllcefion.s. the ft r'nostpreferre$ embodiment is 1,3 millimetres n diameter.

When considering a smaller embryo (such as C Elegarisi the dimensions of the wells 2$ may be adjusted appropriately during manufacture of the receptacles 10.

it. Such an arrangement may be readily manufactured usng the well layer 2$ o the invention. since the remaining parts of the receptacle 10 may be standardised and only the welt Layer 2$ modified in accordance witn the requirement of the precise experiment under consideration, before assembly of the receptacle takes piace.

so Advantages ct selecting the well dimensions specifically for a partoular experiment include: * tess precision is required to inject embryos nto the well; as there is less scattering of light by the culture medium than would otherwise occur; the fluid channels of the kind described herein may be accurately located for provdn nutrients to the cell arid/or removing waste products; aside from the benefit of providnq mirrors and/nt prisms 27. 28 the, well layer'24 additionally may provde for possible differential interference contrast (DIG) microscopy, The foregoing advantages anse in addition to the benefits of good soluble signalling and the provision of adequate nutrients for a pnolonged period of actMty, that are mtpianed hereinabove, ML flC'WS th SCh vtcaUy secioned IeW aoordane with nvetior ard defnd herein fri to:ke standard fltre weL of ftie tvp that +--Spacer member 57 nciudes tomed therein one or more fhrouqhqoinp bores 58 nernetting the transport of nud from one side at the spacer member 57 to the other. This permts he resup lyina at nutrients, etc. to ceils in the cuitivation space.

t; A modified version of the spacer member 57' is shown in schematic. verhcaiiy sectioned VeW'':n flgure 10.

The Ftture 10 spacer nember ST inciudes mod'ihed bores 58 that taper, as iUustrated, in the downward direction. Such bores are suitable for tarqehnp of introduced nuthents tO onto the cebs or other aartioles 23 being cultivated Within the receptacie 10. The use of the trustoconicai or other tapered shapes shown assisis in accurate targeting of inected iiuais, Figure. i 0 aiso iilust,rates somewhat schematicaiiv a further, optional feature of the 1 spacer member 57' nameiy the presence of features sube as teet 59 that engage with cornpiementarv teatures (such as recesses) formed in or secured to the interior of recess The purpose of such engagement is to assure that the spacer member ST is Sways nserteri nto the' recess 1$ at the correct location and in the. right oneniation for taebn ci PuSs or' to the particies 23, Other ibrms of inter"enaaoenient members may of course be provided actinq between the soacer mernoer 5$' and the structure of the recess 12, As a further example the Sacs of the spacer member 57' may be arran ed to engage the iateral 000nc:ary 14 so as to p revent rotation of the spacer member, Yet a further arrangement is shown in Figure 11, in wSch the spa member 57 onits the teet 59 and instead is supported on one or more horizomav extending sho.uiders $2 ac extenchno radiany nwardly from the later'S boundary 14 at the icwer end of recess 12.

Such a means of supporhno the spacer member 57 may be of partcuiar benefit when the receotacie 10 mdudes a weD $5 of the kind described above.

The feet SP are aiso present in the Figure 9 embodiment, but in that case they serve smpiy to rase the spacer member 57 off the recess bottom 15 so as. to de.hne an adequate ecU culuvation space, Fçur O 2$ rs1e on q& ayer 1 s ustrt& ft somo mod f use th reoepice own n Fure 10 the spaoer ye 5T ma be removed after ard.diehno of The ce h; nrr1 Th rr r Fgue $ K)W9 var Ot recptce I 0 wthn tr sco of the rwentbn.

Foure 14 th 12 end the ths member 21 crc eeoh fr tovremda ---" in. such experiments in accordance with the method of the mventon. the recess bottom 1$ may te initially coatec with antibodies that are in some way antiqenic, or at least mutagenc, to the parüdes of interest. Such coatinq may be achiever.'; by DipetUno the anticens onto the recess bottom 1$ edth the closure 18 in be open pobeon.

Thereafter the cicsure 1$ is closed so as to define the fluid flow path 6$. The port 67 may be connected to a fluid supply containino the particles of mterest, or en in ctor 68 employed tO charge the fluid flow path wiTh fluid. As the particles resultinoly flow over the recess bottom 1$ they become attached to the antibodies.

The f)uic; flow path 64 may then be further employed for the purpose ci' flushino ure adhered oar cies away from the cell cultivation space The adhered particles are then available for detection, experimentation andibr study.

Using the arrangement of Figure 14 multiple cell cultivation spaces may be treated usinq a sinr.$e sourc'e.

a refinement of the l'oregoing method the fluid flow path 64 may subsequenflv be employed for the purpose of conveying further antigens to the target particles into the cell cultivation space(s) Such further antigens may be ep radiolabelieri or tagged with.

fluorescent taps. Subsequent scintigraphy or optical detection tests ma be employed to identify the particles, of interest, to which the further antibodies have adhered.

Among other advantages of the foregoing technique are the foiiowinp, * on;y a small area of the recess 1$ is initially coated with antiboches, thereby saving on matenais and energy; at the coated area the cell cultivation space may be eg. only two or three partcles high. leading to good particle adherence.; * the Reynoids number of fluid outside the (narrow) fluid flow path, 64 is reiative hih, fvvinq rise to low pumpnq pressure reouirements and minimal shear forces; with the closure member 1 9 in its open position flushinq with air or water / other solvents are readily possihki; $2 0 iLHd flow can be eflectad in either direction aion the &fld flow path 54, and muttiie fluhi passes are possib'e: * as seted, a sngie fluid source can be used to supoiy fluid th muitiplg, nterconnected recesses.

Claims (5)

1. CLAIMSA ce culuvatlon recep;acie ccmprisnq a recess layer havincl fomierl therein a downwardly extenthny recess deflninp an openno on an upper side of the recess laer S and turther defining one or more ateral bourgianes that taper inwardly in the oownward from the openino, to ntersect the bottom ol the recess; a bottom Layer defining the said bottom; and a removable closure for the opening ncluono a closure member that s moveabie from an open posihon spaced from the aperture to a closed oostticn in which it closes off the aperture the closure. member inciudinq extanthnp downwardly 0 Porn an underside a displacement member that when the closure rnemberadoc.ts its dosed oosbon occupies part of the interior of the nacess, the displacement member rerrmnatinci in a tower surface. that is spaced from the recess bottom to define a ce cultivation space in the recess and the displacement member includinc, at least a portion that tapers in the said downward threction at a more acute taper angle than the taper is anole of the or each said lateral boundary.
2. 2. A receptacle accordin to Claim I wherein the horizontal cnosssection of the displacement member is essentially complementary to the horizontal cross section of the recess at the same depth in the receptacle,
3. 3. A receptacle accordina to Claim I wherem the recess is croular in horizontal crosesection and is frustoconical in shape.
4. 4. A receptacle according to Claim 3 wherein the dsplacement member is cIrcular in honzontal orosssection and s frustoconical in shape.
5. 5. A receptacle according to Claim I wherein the nacess s polygonal in horizontal cross..section a is frustopyramidal in shape u. A receptacle according to Claim 5 wherein the thaplacement member is polygonal In horlzontal crcssse.ction and is frustopyramdal in shape.7. A rec.eptacie according to any' preceding claim wherein the or a said ie,teral boundary subiends an angle. of between 170 degrees and 100 degrees, and especially S 1Sf) degrees to 110 degrees, to the bottom of the. recess.S. A recepthce accorcnnq to any preceding claim wherein the bottom of the recess cOnCaV&y curved.9. A receptade accordiop to any precedin cm, wherein the lateral boundary is conceiy curved.10, A receptacie accordinq to Cam 9 wherein the thee of the iaterai boundary is cars boUc when viewed in verti section.11 A receotacie eccordinu to Cisim 8 wherein the or a saul iaterai bouno'anv subtends sri angle of between I 70 degrees and 100 degrees. and especiafly 150 degrees to 110 degrees. to a tancierit to the bottom c4 the recess.12. A receptacle accorthnc to any of Claims B to II wherein the lower surface of the o'isplacement member is convey or concavely curved.13, A receptacle according to any preceding claim wherein the recess layer is transparent, and is supported on a base layer.14. A receptacle aocordinct to Claim 13 whereuri the base layer is transparent.15, A receptacle according to Claim 13 wherein the recess layer w integrai with the base leer.15. A receptacle according to ClaIm 13 wherein the recess layer is formed separately from the base leer 17. A receptacle according to Claim 16 wherein the recess layer and the base layer are r easabiv securabie one to the other, IS. A receptacle accordinc to any precedinci claim wherein the closure member and the dispacement nienoer are. tcrmeo rrom one or more transparent rnarenais..19. A receptacle according to Claim 13 wherein the closure member and the displacement rr'ernber are integral with one another.20. A receptacle according to any preceding claim inciudng a plurality of the recesses forriad in the recess layer.$1. A recentacla accordino Ic Clam 20 rciudinct a respective said displacement S member for each said recess 22. A, recepaceaccorthro to Claim 21 including a common closure member havrnq extending downwardly from an underside a plurality of displacement members each kca.ted so as to berletate a said recess on. movng of the closure member to its closed tO posihon.23. A receptacle accordin to Claim 21 inciuthng a piurahty of closure members each hain one or more displacement members axtending downwardiv from an underagie, s 24. A receptacle according to any of Ciams 20 to 23 including one or more microchanneis interconnectinc two or more of the' recesses, 25. A receptacle according to Claim 24 when dependent from Claim 1$ or any precethng claim iependng therefrom, wherein the or each mmroc,hannel is defined at the 2!) boundary of the recess layer and the base layer.$6. A receptacle according to Clam 24 or Claim 25 inciudinq defined in the closure member an observation region at which optical signals generated in the nuicrochannetare. detectable.27. *\.rc ceptacie according to any of Claims 24 to 26 ncluding one or more elements selected from the list comprising optical sensors, chemical sensors and flow controi elements located so as to. act or. or be influenced by fliad in a said fluidic micro-channel.23. A receptacle accordinc. to any of Claims $4 to 27 including an electroosmosis element extending-from the exterior of a said receptacJe to the interior of a said miorochannel and being connected-to a voltage source.$9. A receptacle, according to. any of Claims 20 to 28, wherem, the plurality ty* recesses are arranged in a regular pattern in the recess layer'.30. A receptacle accordlln to Ciaim 29 wherein, the plurality of recesses are arranqed n mutualyort oçgsnai rows and coumns so as to detne a grithilke array.31. A receptacle according to any preceding dairn further ncfudtng a wsll unoeriymp' the or a sac recess and in reqister therewltft 3$. A reepfcle acoordinq to Ciam $1 including a well iaer underlying the recess layer and ha\'lnq formed therein the well.ic 33. A receptacle according to Claim 32 whereir' the well layer is transoarent.3$. A rereptacte accordina to Claim 32 wherein the well layer inchides formed or o'efined therem one or more optical pnsrns and/or rectors'.:35, A receptacle accordmo to any of Claims 31 to 34 when dependent from Cialm 20, noludin a reapedllve said well underlying each saci recess.36. A receptacle according to any precethng claim, wherein the displacemert member includes formed therein one or more flue; flow oassacle9 that penmit the conveyance of fluid to and/or from the Interior ot the recess.37. A receptacle. according to Claim 36 wherein the or each said passaqe terrnriates nafurrher operano in the said lower surface.35. A receptacle according to Otaim 37 indhxling a plurality of hne passages and a plurellty of the further openings that are arranged in a. regular or rrequiar patternn the lower surface.39. A receptacle acooiding to any of Clams 36 to 38 wherein the or each Passage w connectahie to a source and/or a draIn of fluid.A receptacle accordIng to any of Claims 36 to 39 incL:ing a pluralit of thsplacement members two or more of which each include a saId passage and are fluidica liv connected to one another.41. A receptacle acccrdinç to Claim 31 or ens orecedino claIm Oet3eno'no therefrom wherenn the displacement member substantially occupies the recess when Pa closure closes off the opening.o 42. A ceptac.le according to any precedin claim induthno received w'Thn the recess one or mom spacer members that space the displacement member from the bottom o the rec.eso.43. A receptacle according to Claim $2 including in The recess a fluid of lower density Then the matehal of the spacer member.$4. A receptacle accordion to Claim 42 or Claim 43 wherein the spacer member is formed from a transparent rnatenal.45. A receptacle accorcilnq ro any of Claims 42 to $4, wherein the spacer member includes, tormed Weren one or more throughgcing holes permittinc the passape of flowadie matter fmm one side of the spacer to another 4$. A receptacle accordine to Ciaim $5 wherein one or more: said throuch.qoino holes is fr.ri in shape.47. A receptacle accordLng to any of Claims 42 to 4$ wheren the spacer member includes dependino downwardly therefrom one or more suopo:rt feet defininty a Ca between the unner member and the bottom of the recess.$8. A receptacle according to any of Claims 42 to $7 wheren the spacer member m remc'vabiy osItionabie in the recess.$9. A receptacle accordino to Claim $8 wherem the spacer member and the recess each include one or more mutualiy engageable posillonino merhbers for locating the spacer member in a predetermned location end/or onentation in the recess.50, A receptacle accordion to any preceding claim whereEn the recess is trustopvrarnidai in horizontal crosssectjon anc: the displacement member is also so frustoryvramidai n horizontal crcsssecbon, the displacement member when the closure: adorns a dosed configuration on two opposite sides enoaoing and sealinc against respective. mntnuauy opposed lateral bounoanes ot the recess so as to oetme on two further, muuaHv opposed lateral boundanes a paft of fluId flow reccuis that are interconnected by the. oeU cultIvation space whereby to define a fluid flow path extendinp from one side of the recess to another, the f1ud flow oath being conneotebie to one or more cIevces for effecting flow of fluid in the path 51. A receptacle aocordlnq to Claim 50 includinq one or mere fluid flow corts that are fkcaHy connectable for supplyino fluid to and/or from the fluid flow path.5$. A closure for a ceH oulfivatior; receptacle, including a closure member having In extendincl downwardly from an inuse underside a displacement member that when the olosure adopts a closed position closing off a ceh cultivation recess oocupies part of the Intenor ol the reoess, the dsplacernenf member terminatinp in a lower surface and incdinc at least a portion that tapers in the said downward direction. 53 A closure acoording to Claim 52 that is circular in horizontal cross*-section and frustoconicai in shape.5$. A closure accorthno to Claim 52 that rs polygonal in horizontal cross'-section and frusto pyramidal in shape 55. Acosure according to any of Clams 5$ to 54 wherein: the lower surface. of the displacement nmrnber is conVexly or concavely ourved, 55. A closure according to any of Clams 52 to 55 that is formed from one or more transparent materials, 57 A closure according to any of Claims 5$ to 55 wherein the. closure member and the disr;iace,ment member are. formed integrally with one another.to 58, A closure according to any of Claims 52 to 57 wherein the closure member include.s extending downwardly therefrom a plurality of displacement members, 3$. A closure according to any of Cairns 52 to 53 wherran the displacement member includes formed rheren one or more flwd flow Passaoes, 50. A closure according to Claim 59 wherein the or each said passage terminates in a further opening m the said lower surface. 3$61 A closure a000rthno to Ctatrn 60 idudino a pwraftty of the passages and a Dturaht of the further Deenmas that are afranged n a ns utar or frrequiar pattern in the lower surface.62. A dosure according to any ol Cirns 59 to $1 wherein the or each: passage is connectabie. to a source and/or a drain of fluid.63. A ciosure accorthng to Cirn 58 or any preceding claim depenthng therefrom, O ncluthnq a piurality of the said passages that are I uidioaliy mutuady interconnected, 6$. A ccli culture apparatus including one or more receptacies accortang to any of Ciaims to 5$.$5. An apparatus accorthng to Claim 64 including one or more ciamps for securing the or each receptacle relative lo the apparatus.66. An, apparatus accordinc4 to Claim 65 wherein the damp is or'nciuc1es a Perforated suc6on piate that is operahv&y connected to a Suction pap. An apparatus accordin to any of Cairns 64 to fib includinoen niector havng one or more cannuiae for perforating the closure member so as to permit injection of matter nto the interior of the or a said recess through the closure merntar, a 6'S. An, apparatus accorc:in to Cairn 67 wherein the injector includes two cannulae, that respectvey inject material ntr the intenor of the recess and extract materiai theretrorn.$0 An apparatus accordinq to Claim 68 wherein the cannulae are supported on a ac comrnop support that is moveabie relative to at least one said recess sc as toperniit nsention of the cannulae into and withdrawal of the cannulae from the said recess:.70. An apparatus acccrthno to Claim $9 wherein the support supports a plurality of niectors arranoed in a pattern corresponding' to the pattern of a piurahty of the recesses. $071 An arparatus accoiiing to any of ClaIms 67 to 70 when decending from Claim fit) or Claim 51 wherein one or more said cannuiae in use is fluidicaily connectable to th fluid flow path.72. A method 01 treatinc parficles comprising the steps of: coating the recess bottom of a recepiacle accordinq to either of Claims 51 to 52 or of a receptacle of an apparatus accordino to Claim 71, having ts closure memper in an coen position. wth antibods; (id causing the closure member to adopt its closed position thereby defimno the 11(50 flow path; (iii) causing a flrst flud, containinp one or more particles to which the said antibodies. are antigenic, to flow in the fluid flow path; (ni) causinc a seconc, rinsina fluid to flow in the fluid flow petit and (v) detecting the particles in the recess.$3. A method according to Clami 72 ncluding the further steps oh (vi) causing a tftrd flud.. containing optically detectable. kirther antiboaies, that are antiqenc to the particles to flow in the Ilud flow path; and (vii) detecting the particies usint; one or more optical de.fecticn technipues.7$. A recentacle oeneraliy as herein described, wth reference, to and/or as iliustraleci ifl the aocomnpar:vinq crawings.75. A closure generafly as herein described, with reference to and/or as illustrated in the accornpanyino drawinps.7$. An apoaratus generafly as heren described, with reference to andor as illustrated in the accompanying drawings.77. A method generally as herein described, with reference to and/or as illustrated in the accornoansno drawings. $1