CN1348103A - Planar microelectromagnetic unit array chip capable of being gated separately - Google Patents

Planar microelectromagnetic unit array chip capable of being gated separately Download PDF

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Publication number
CN1348103A
CN1348103A CN 01136350 CN01136350A CN1348103A CN 1348103 A CN1348103 A CN 1348103A CN 01136350 CN01136350 CN 01136350 CN 01136350 A CN01136350 A CN 01136350A CN 1348103 A CN1348103 A CN 1348103A
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China
Prior art keywords
chip
electromagnetic
magnetic
little
molecule
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CN 01136350
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Chinese (zh)
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CN100476431C (en
Inventor
吴镭
王小波
程京
杨卫平
周玉祥
刘理天
许俊泉
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BOAO BIOCHIP Co Ltd BEIJING
Tsinghua University
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BOAO BIOCHIP Co Ltd BEIJING
Tsinghua University
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Priority claimed from US09/685,410 external-priority patent/US6716642B1/en
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Abstract

The present invention discloses array electromagnetic chip and electromagnetic biochip comprising microelectromagnetic units capable of being gated separately and the method of utilizing these chips to control biomolecule, chemical reagent and other particles and microstructure directionally.

Description

Little electromagnetic units array chip of the planar structure of gating separately
The present invention is Chinese patent application " electromagnetic units array chip but the single-point gating declines; electromagnetic biological chip and application " (number of patent application: 99104113.5, submit the date: on March 15th, 1999), U.S. Patent application " Individually Addressable Micro-Electromagnetic Unit ArrayChips " (Patent Office's numbering: 09/399,299, submit the date: on September 17th, 1999) and international patent application " Individually Addressable Micro-Electromagnetic Unit Array Chips " (Patent Office numbering: PCT/US99/21417 submits the date: extendible portion on September 17th, 1999).
Also quoted following patented claim among the present invention:
International patent application " Method for Manipulating Moieties in MicrofluidicSystems " (Patent Office's numbering: PCT/US00/25381, submit the date: on September 15th, 2000, inventor: Xiaobo Wang, Lei Wu, Jing Cheng, Weiping Yand, JunquanXu).
U.S. Patent application " Apparatus for Switching and Manipulating Particles andMethod of Use Thereof " (Patent Office's numbering: 09/678,263, act on behalf of office's numbering: ARTLNCO.002A, submit the date: on October 3rd, 2000, inventor: Xiaobo Wang, Weiping Yang, JunQuan Xu, Jing Cheng Lei Wu) and its pairing Chinese patent application " be used for particulate and handle device and using method thereof with the particulate guiding " (Patent Office's numbering: 00129043.6, act on behalf of office's numbering: 12000723EB, submit the date: on September 27th, 2000).
U.S. Patent application " Apparatuses Containing Multiple Active ForceGenerating Elements and Uses Thereof " (Patent Office's numbering: 09/679,024, act on behalf of office's numbering: 471842000400, submit the date: on October 4th, 2000, inventor: XiaoboWang, Jing Cheng, Lei Wu, JunQuan Xu, Weiping Yang) and its pairing Chinese patent application " multi-force operator and application thereof " (Patent Office numbering: 00130563.8, act on behalf of office's numbering: 12000725EB, submit the date: on September 30th, 2000).
U.S. Patent application " Methods for Manipulating Moieties in MicrofluidicSystems " (Patent Office's numbering: 09/636,104, submit the date: on August 10th, 2000).
U.S. Patent application " Methods and Compositions for Identifying Nucleic AcidMolecules Using Nucleolytic Activities and Hybridization " (Patent Office's numbering: 09/684,081, submit the date: on August 25th, 2000).
U.S. Patent application " Compositions and Methods for Separation of Moieties onChips " (Patent Office's numbering: 09/686,737, act on behalf of office's numbering: ART-00102.P.1, submit the date: on October 10th, 2000, inventor: JunQuan Xu, Xiaobo Wang, Jing Cheng, Weiping Yang, Lei Wu) and corresponding Chinese patent " method of entity molecule separating and required device and reagent on the chip " (Patent Office's numbering: 00131649.4, act on behalf of office's numbering: 12000726EB, submit the date: on October 9th, 2000).
U.S. Patent application " An Integrated Biochip System for Sample Preparationand Analysis " (Patent Office's numbering: 60/239,299, act on behalf of office's numbering: ART-00102.R1, submit the date: on October 10th, 2000, inventor: Jing Cheng etc.).
Technical field
The present invention relates to use micromachining technology method that make, that utilize magnetic field force to carry out the biochip of chemistry, biology or biochemical reaction and analysis and use this class biochip to carry out chemistry, biology or biochemical reaction and analysis.
Background technology
Biochip technology is the emerging technology in life science and biomedical research field in recent years, application at biology, biotechnology and biomedical sector is very extensive, for example point mutation detection, dna sequencing, gene expression analysis, drug screening and clinical diagnosis.Biochip is the micro device that a class can be used for chemistry or biological respinse, utilize the microelectronics of semi-conductor industry and the similar pass of micro-processing technology art to make with other, can be with existing discrete chemistry or biochemical analysis process and device integrated and be simplified in the device based on microchip.A large amount of technical papers all relates to this class device now.
By following article, can see the extensive use of biochip." Rapid determination ofsingle base mismatch mutation in DNA hybrids by direct electric fieldcontrol ", Sosnowski, (Proceedings of the National Academy ofSciences such as R.G., USA, 94:1119-1123,1997), " Large-scale identification; mapping and genotyping of single-nucleotide polymorphisms in the humangenome ", Wang, D.G. etc. (Science, 280:1077-1082,1998) have introduced the application of biochip on point mutation detects; " Accurate sequencing by hybridization forDNA diagnostics and individual genomics ", Dumanac, (NatureBiotechnology such as S., 16:54-58,1998), " Quantitative phenotypic analysis ofyeast mutants using a highly parallel molecular bar-coding strategy ", Shoemake r, (Nature Genetics such as D.D., 14:450-456,1996), and " Accessing genetic information with high density DNA arrays. ", Chee, (Science such as M., 274:610-614,1996) introduced the application of biochip technology aspect dna sequencing; The application of biochip in the gene expression monitoring can be seen in following article: " Genome-wide expression monitoring in Saccharomyces cerevisiae. ", Wodicka, (Nature Biotechnology such as L., 15:1359-1367,1997), " Genomicsand human disease-variations on variation. ", Brown, P.O., Hartwell, L., " Towards Arabidopsis genome analysis:monitoring expression profilesof 1400 genes using cDNA microarrays. ", Ruan, Y etc. (The Plant Journal15:821-833,1998); " Selecting effective antisense reagents oncombinatorial oligonucleotide arrays. ", Milner, (NatureBiotechnology such as N., 15:537-541,1997), " Drug target validation andidentification of secondary drug target effects using DNA microarray. ", Marton, (Nature Medicine such as M.J., 4:1293-1301,1998) introduced the application of biochip aspect drug screening." Cystic fibrosis mutation detection by hybridizationto light-generated DNA probe arrays. ", Cronin, (Human Mutation such as M.T, 7:244-255,1996), " Polypyrrole DNA chip on a silicon device:Example ofhepatitis C virus genotyping. ", Livache, T. etc. (Analytical Biochemistry.255:188-194,1998) have introduced the application of biochip in clinical diagnosis.By on can see the widespread use of biochip.
It is to make by biomolecule (for example, oligonucleotides, cDNA and antibody) is fixed on chip surface that one class biochip is arranged.There are many kinds of methods can be used to make this class biochip.For example United States Patent (USP) 5 (is seen by Affymetrix company, 445,934 and 5,856,174) developed a kind of method of making biochip by light guiding chemosynthesis, the biomolecule of chip surface is synthetic by the synthetic method of solid phase photochemistry by the photolithography technology.The method that a kind of spray regime is made biochip has been invented by Incyte Pharmaceutical company (seeing United States Patent (USP) 5,874,554), and previously prepared cDNA is positioned at the surface of chip by the mode of direct injection.Thereby the contact point sample method of Stanford university invention is to drive the sample dispensing head by high speed, high accuracy mechanical arm directly to contact the surface (seeing Schena, M.et al.Science 270:467-70 (1995)) that sample is arranged in chip with chip.The making that a kind of mononucleotide probe synthetic method is used for biochip has been invented by the Washington university that is positioned at Seattle (Seattle), four different piezoelectricity dispensing heads are respectively charged into four kinds of different nucleotide, distribute different nucleotide sequentially by controlling these piezoelectricity dispensing heads, thereby be implemented in the automatic synthetic oligonucleotide of chip surface and (see Blanchard, A.P.et al.Biosensors ﹠amp; Bioelectronics 11:687-90 (1996)).Hyseq company has developed the passive type membrane unit (seeing United States Patent (USP) 5,695,940 and 5,525,464) that is used for gene order-checking.
Biochip mainly is divided into two big classes: active and passive type.The passive type biochip is meant that the chemistry that takes place on those chips or biochemical reaction are that passive diffusion by sample molecule realizes.And in active biochip, reactant can be transported and enrichment by applied external force, and reaction is just carried out under the acting in conjunction of the diffusion of reactant suffered applied external force on it itself like this.Present most obtainable biochips are for example from the oligonucleotide DNA chip of Affymetrix company with all belong to the category of passive type biochip from the cDNA chip of Incyte Pharmaceutical company.Active structurally closely similar with the passive type biochip, all on chip, fix the array that constitutes by different aglucons.By using different marks, detectable mark, detection system and indication molecule (for example luminescent dye molecule), the reaction between aglucon and the corresponding molecule can be able to qualitative or quantitative detection.Like this, the biochip that contains the array of being made up of the various fixed aglucon can carry out the detection of multiple material simultaneously.
Little processing and microelectric technique are not fully used in the design of existing many passive type biochips.And passive type biochip and being not suitable for the integrated micro-total analysis system that advances to realize from initial specimen preparation to final Molecular Detection.In addition, the passive type biochip also exists other shortcoming, and for example sensitivity for analysis is still not high enough, the reaction time is long, be difficult to realize that temperature, pressure, electric field and the molecular conecentration in the single zone (can be called the unit) to chip surface effectively control or the like.
On the contrary, active biochip carries out microfluidic manipulations or electricity manipulation by reactant being applied applied external force, realizes manipulation, reaction (for example PCR), the hybridization to molecule and separates (for example Capillary Electrophoresis).But simultaneously, many so active biochips do not have high-throughout parallel processing capability again.The electronics biochip of Nanogen company development can be handled and control sample biomolecule by the electric field that microelectrode produces, and compares with the passive type biochip, has improved the sensitivity of reaction velocity and detection significantly and (has seen United States Patent (USP) 5,605,662,5,632,957 and 5,849,486).But in order in solution or suspension, to utilize the effective mobile biomolecule of electric field, the conductivity of solution must be lower, this has limited the selection of biochemical analysis damping fluid greatly, many enzymes and other biomolecule be sex change easily in such low ionic strength environment, and easily and chip surface non-specific absorption takes place.
The present invention has provided a kind of design of new active biochip, wherein applied external force is a magnetic force, is by the cell array of gating generation separately on the chip.In this design, magnetic force can be used to control and handle molecule or the particle that magnetic is modified, thereby promotes the interaction of molecules of chip surface.Magnetic force has been widely used in biology, chemistry and biochemical field.For example, in the method for magnetic cell sorting, pan coating can special and corresponding cell combination in cell mixture at the magnetic-particle of the antibody of specific cell, in conjunction with after, cell-magnetic-particle compound can optionally transport under magneticaction, thereby the sorting that realizes magnetic force (is seen Miltenyi, S.et al.AHighgradient magnetic cell-separation with MACS (Cytometry 11:231-236 (1990)).In addition, at United States Patent (USP) 5,439,586 have provided a kind of design of magnetic filter of three-dimensional, can be used for separating magnetic-particle and non-magnetic particle, United States Patent (USP) 5 at fluid, 655,665 have provided a kind of design of micromechanics magnetic-particle separation vessel, can be used for microfluid magnetic and separate.
Description of drawings
Fig. 1 is the structural drawing of the little electromagnetic units array chip among the present invention, though drawn electromagnetic unit is a spatial structure among the figure, electromagnetic unit also can only be made planar structure;
Fig. 2 shows in little electromagnetic units array chip among the present invention that the unit that is applied electric signal by gating attracts the phenomenon of magnetisable material, though drawn electromagnetic unit is a spatial structure among the figure, electromagnetic unit also can only be made planar structure;
Fig. 3 is the structural drawing of the little electromagnetic units array chip among the present invention, though drawn electromagnetic unit is a spatial structure among the figure, electromagnetic unit also can only be made planar structure;
Fig. 4 is the sectional view of chip shown in Figure 5, though drawn electromagnetic unit is a spatial structure among the figure, electromagnetic unit also can only be made planar structure;
Fig. 5 is the structural drawing of the little electromagnetic units array chip among the present invention, but wherein little electromagnetic unit of single-point gating constitutes a square formation, and what provide among the figure is vertical view.Though drawn electromagnetic unit is a spatial structure among the figure, electromagnetic unit also can only be made planar structure;
Fig. 6 is the structural drawing of the little electromagnetic unit among the present invention, thereby the gating of unit (for example generates the magnetic field realization by applying electric current, after applying electric current, magnetic core is magnetized, produce magnetic field), though drawn electromagnetic unit is a spatial structure among the figure, electromagnetic unit also can only be made planar structure;
Fig. 7 is the synoptic diagram of first group of lead that is used for forming the miniature coils of wound core among the present invention, though drawn among the figure be spatial structure, electromagnetic unit also can only be made planar structure;
Fig. 8 is the synoptic diagram of second group of lead that is used for forming the miniature coils of wound core among the present invention, though drawn among the figure be spatial structure, electromagnetic unit also can only be made planar structure;
Fig. 9 is the synoptic diagram of the 3rd group of lead that is used for forming the miniature coils of wound core among the present invention, though drawn among the figure be spatial structure, electromagnetic unit also can only be made planar structure;
Figure 10 is the schematic diagram that each little electromagnetic unit is carried out independent gating by electric switch, each unit is that the electric switch by two series connection is connected with power supply and ground wire, the break-make of these two electric switches is to control by the electric signal that is applied on the ranks lead, though drawn among the figure is spatial structure, electromagnetic unit also can only be made planar structure;
Electronic switch shown in Figure 11 A can be a bipolar transistor, though drawn among the figure be spatial structure, electromagnetic unit also can only be made planar structure;
Electronic switch shown in Figure 11 B can be a metal oxide semiconductor field effect tube (MOSFET), though drawn among the figure be spatial structure, electromagnetic unit also can only be made planar structure;
Figure 11 C is the circuit diagram in the lcd technology used in the present invention;
The structural representation that Figure 12 is the current biochip of inventing after installing the window that a fluid pool and is used for optical detection, though drawn among the figure be spatial structure, electromagnetic unit also can only be made planar structure;
Figure 13 schematic representation aglucon or target sample molecule carried out magnetic modify by cutting chemical linking arm, though drawn among the figure be spatial structure, electromagnetic unit also can only be made planar structure;
Figure 14 A is the vertical view of the electromagnetic unit of midplane structure of the present invention, is magnetic core (100) and coil (110) among the figure; Figure 14 B, Figure 14 C and Figure 14 D are respectively the photos of little electromagnetic unit of the present invention;
Figure 15 is the logarithm equal-value map in the magnetic field of magnetic core in vertical direction 200 microns * 200 micron planars, and the coordinate of figure begins to count from the center of magnetic core, is of a size of 200 microns * 200 microns * 5 microns magnetic core;
Figure 16 is the logarithmic graph with the magnetic field force of Figure 15 in a plane;
Figure 17 is the chip figure with 1 centimetre of size of 1 cm x of 16 horizontal cells;
Figure 18 then is the silicon chip figure that is manufactured with 16 chip blocks on Figure 17;
Figure 19 A be among Figure 14 A along the sectional view of A-A direction, Figure 19 B be among Figure 14 A along the sectional view of B-B direction, be magnetic core (100), coil (110), megohmite insulant (120), coating (130) and conductor material (140) among the figure;
Figure 20 is the calibration curve of the MR head in the electromagnetic unit of the present invention,
Figure 21 is the field intensity-distance map that is of a size of 400 * 50 * 5 magnetic core;
Figure 22 is the field intensity-map of current that is of a size of 400 * 50 * 5 magnetic core;
Figure 23 is of a size of on 1600 * 50 * 5 the magnetic core to apply 20 milliamperes electric current, removes the remanent magnetism curve of this process of electric current then suddenly; Right figure among Figure 23 applies 20 milliamperes of electric currents on the device, the amplitude with electric current decays according to sinusoidal manner then, until the remanent magnetism curve that drops to zero this process;
Figure 24 A and Figure 24 B are two kinds of design drawings of chip of the present invention, Figure 24 A is one and has 16 chips of the electromagnetic unit of gating separately, Figure 24 B has two groups each 8 electromagnetic units of gating separately, and the chip among Figure 24 B is the first-selected structure of the capable ripple of magnetic force of the present invention;
Figure 25 is the figure that is shaped on the silicon chip of a plurality of chips among the present invention, and wherein the size of silicon chip is generally 3 inches;
Figure 26 is the structural drawing of the horizontal cell with taper magnetic core end structure in the groove that is produced on chip surface of the present invention;
Figure 27 has provided the structural drawing of multiple magnetic core end (160) and groove (150);
Figure 28 is the target molecule that is connected on the magnetic particle;
The synoptic diagram of Figure 29 for the freezing particulate that comprises aglucon molecule and magnetic particle being picked up with the magnetic divider;
Figure 30 is for being discharged into the freezing particulate among Figure 29 the synoptic diagram of the biochip surface among the present invention; Though drawn among the figure is spatial structure, electromagnetic unit also can only be made planar structure;
Figure 31 represents that the freezing particulate that will comprise aglucon molecule and magnetic particle among Figure 30 melts; Though drawn among the figure is spatial structure, electromagnetic unit also can only be made planar structure;
Figure 32 represents magnetic particle among Figure 31 and aglucon molecular separation; Though drawn among the figure is spatial structure, electromagnetic unit also can only be made planar structure;
Figure 33 represents to do random motion through the target sample molecule that magnetic is modified in institute's invention biochip surface; Though drawn among the figure is spatial structure, electromagnetic unit also can only be made planar structure;
Figure 34 signal is flow through on the electromagnetic chip multiple conducting wires to encourage the electric current type of flow synoptic diagram of one group of little electromagnetic unit (even one group of magnetic core magnetization) by certain way; Though drawn among the figure is spatial structure, electromagnetic unit also can only be made planar structure;
Figure 35 represents to flow through the another kind of different current-mode of electromagnetic chip lead, this electric current type of flow can encourage the electric current type of flow shown in Figure 34 little electromagnetic unit that can not encourage; Though drawn among the figure is spatial structure, electromagnetic unit also can only be made planar structure;
Target sample molecule after Figure 36 represents to modify through magnetic and be fixed on the chip aglucon in conjunction with after, thereby be fixed on the electromagnetic chip surface; Though drawn among the figure is spatial structure, electromagnetic unit also can only be made planar structure;
Figure 37 is the principle schematic of after the aglucon of biochip surface among target sample molecule and the present invention reacts magnetic particle being excised from target sample molecule; Though drawn among the figure is spatial structure, electromagnetic unit also can only be made planar structure;
Use the capable ripple magnetophoresis device (170) that has little electromagnetic unit (190) of the present invention that magnetic particle (180) is gone the ripple magnetophoresis among Figure 38; Figure 38 A, Figure 38 B and Figure 38 C shown at little electromagnetic unit gating and when closing magnetic particle along the situation of the capable wave motion of magnetic force;
Figure 39 has provided the particulate switch figure that uses in the present invention's row ripple magnetophoresis; Figure 39 A is switch (200); Figure 39 B is the direct current that produces the capable ripple of magnetic force; Figure 39 C is the alternating current that produces the capable ripple of magnetic force; Figure 39 D is the sinusoidal ac that produces the capable ripple of magnetic force;
Figure 40 is that the present invention detects the method that cell (for example lymphocyte) distributes, and first-selected method is the cell magnetophoresis;
Brief summary of the invention
The present invention recognizes, in order to realize biology, chemistry, machinery and physical process or automation of operation and microminiaturization, must effectively handle entity molecules such as biomolecule or particle.Especially, method that can special and accurate controlled entity molecule position is very useful in the occasion that needs detect the material of combination on entity molecule or the entity molecule.
The present invention has designed a kind of electromagnetic biological chip, but little electromagnetic electrode unit of single-point gating is arranged on the chip, and one or a plurality of array are formed in these unit.Can have one or a plurality of little electromagnetic units array on the electromagnetic biological chip.Thereby each unit can apply electric signal separately and produce magnetic field.The magnetic field that each unit produces can or be closed by independent gating, and the intensity in magnetic field and phase place also can be regulated according to the exchange current that is applied on the unit.The magnetic field that chip surface produces can be used to handle the material with magnetic, comprises the compound that magnetic-particle, biomolecule, cell, other particles or these materials and magnetic-particle are formed.Material with magnetic can be under the ordering about in magnetic field, to the motion of the precalculated position of chip surface.The chemical modification that the surface of chip or the part of chip surface can be carried out, thus the fixedly functional layer of aglucon molecule formed, so that the material and the aglucon molecule generation compatible reaction of magnetic force guiding.Increase the speed of different biochemistry and chemical reaction thereby the magnetic guiding has changed the reactant local concentration with manipulation, thereby reached the purpose that improves sensitivity for analysis.Because ionic strength and damping fluid characteristic are very little to influence of magnetic field, even do not act on, so just can select to help most the damping fluid of biochemical reaction.And, do not exist can cause electrochemical reaction and produce and make the harmful effect of analyzing or reacting complicated highfield.
The present invention has further disclosed the method for handling magnetic particle on electromagnetic chip.Magnetic particle can be suspended in certain fluid (aqueous solution or non-aqueous solution) or in air, even in a vacuum.When a little electromagnetic unit was passed to electric current, near the magnetic particle the unit can be subjected to the effect of magnetic force, was adsorbed to the surface of this unit.Promptly when magnetic particle suspending liquid was covered with entire chip, unit of gating only can influence the magnetic particle with this unit direct neighbor.But if, just might move and accumulate in the magnetic particle that all are suspending on the entire chip by giving each unit energising in an orderly manner.Such cooperative motion is called " manipulation ", and this manipulation can be controlled by open or turn-off the unit by predefined procedure.Manipulation comprise that also being added in direction of current on little electromagnetic unit by change changes Distribution of Magnetic Field and act on power on the magnetic particle, reach and change and the position of control magnetic particle the purpose of speed and other motion features.According to different request for utilizations, can allow units synchronization all or part switch on.Certainly also can switch on one by one to little electromagnetic unit.
The present invention has further disclosed and has utilized electromagnetic chip to handle biomolecule/biological particle, chemical reagent molecule, the method for drug molecule or other forms of molecule, particulate.Generally speaking, electromagnetic chip can be used to handle any magnetic particle.In order to control and handle non-magnetic particles/molecule, need carry out magnetic to them and modify.For example, molecule can be connected to the magnetic particle surface by covalently bound or physisorption.These biomolecule can be protein (as antibody, antigen, acceptors), and nucleic acid (as single stranded DNA or RNA) or other molecules are as lipid and carbohydrates.Can modify the electromagnetic chip surface so that fixing can with the interactional aglucon molecule of magnetic particle surface molecular.On the ad-hoc location that is fixed with suitable aglucon molecule, assemble just can impel between them at magnetic field force effect magnetic particulate and interact.The detailed description of the Invention definition
Unless illustrate especially, otherwise the meaning of the scientific and technical terminology of all uses consistent with common sense all among the present invention.The direction of mentioning in the invention such as upper and lower provides at employed device.Some terms and experimental procedure involved in the invention all are very common in this area.But consider that different documents is using these terms or can have certain difference each other when describing these steps, so hereinafter can term and the experimental procedure that be used among the present invention be defined and illustrate.
" magnetic force " is meant that particulate causes the acting force that is subjected to owing to magnetic field.Usually, in order to realize handling the purpose of particulate, particulate should itself have magnetic or can be magnetized.With a magnetic particle of being made by the superparamagnetism material is example, and when particulate is in the magnetic field B, particulate itself can be brought out generation magnetic dipole Φ
Figure A0113635000181
=V p(∏ p-∏ m) H mV wherein pRepresent the volume of particulate, χ pAnd χ mIt is respectively the polarizability of particulate and surrounding medium.Φ mBe the magnetic spectroscopy susceptibility of medium, H mRepresent magnetic field intensity.Be shown below the magnetic force F that particulate is subjected to MagneticThe polarizability and the magnetic field gradient that depend on magnetic dipole: Fmagnetic=0.5V p(χ p-χ m) H mΛ B m, wherein " X " and " Λ " represents dot product and gradient computing respectively.Whether particulate is subjected to the difference that magnetic force depends on the polarizability of particulate and its surrounding medium.Usually particle suspension (polarizability is close to zero) in the non-magnetic media of liquid state like this in order to produce enough magnetic force, just needs to use magnetic particle (polarizability substantially exceeds zero).Magnetic particle speed ν under magnetic force and viscous resistance effect during balance ParticleFor ν particle = F magnetic 6 π η m R is the magnetic particle diameter in the formula, η mCoefficient of viscosity for surrounding medium.
" entity molecule " among the present invention is meant the material that can be handled by electromagnetic force, also can be the material that is had magnetic by cloth own, biological example molecule etc. and the compound that can be connected to form by the material that magnetic field is handled.Entity molecule can be a solid, comprises the solid of suspension, also can exist with dissolved state.Entity molecule can be a molecule, the molecule that can be handled comprises, but be not limited only to, inorganic molecule (comprising ion and mineral compound), can be organic molecule also, comprise amino acid, peptide, protein, glycoprotein, lipoprotein, GLP, fat, fat, sterol, sugar, carbohydrates and other micromolecule or complicated organic molecule.Entity molecule also can be a molecular complex, such as organelle, one or more cells (comprising protokaryon and eukaryotic), pathogen (part that comprises virus, parasite, prion or these materials); Entity molecule also can be crystal, mineral matter, colloid, fragment or the like, can be made up of one or more dead matter such as polymkeric substance, metal, mineral, glass, pottery or the like; Entity molecule also can be molecule aggregate, compound, cell, organelle, virus, pathogen, crystal, colloid or other fragment.Cell can be the cell of any kind of, comprises protokaryon and eukaryotic.Wherein eukaryotic also can be an any kind.Common interested cell comprises, but is not limited only to, leucocyte, malignant cell, stem cell, the early stage cell of differentiation, fetal cell, by the cell of pathogenic infection, bacterial cell or the like.
" entity molecule in the cell " is meant and is positioned at intracellular entity molecule, promptly be arranged in tenuigenin or organelle substrate, on any cell inner membrance, be positioned on the plasma membrane (if existence), or be positioned at cell surface, promptly on the outside surface attached to cytoplasma membrane or cell membrane (if existence).
" aglucon " or " aglucon molecule " be meant those can with the biology/chemical molecular of other molecular reaction/combination.For example, aglucon can be a nucleic acid molecules, it can with the nucleic acid chains generation hybridization reaction of complementation.Aglucon also can be an antibody molecule, it can with the antigen or the epi-position combination of correspondence.Aglucon also can be the receptor binding domain or the part of this land.In the present invention, the surface particulate that is connected with above-mentioned aglucon also belongs to the category of aglucon.
" array " is that little electromagnetic unit forms according to certain form arrangement on the chip.Can include several separate arrays on the chip.For example, the array that forms of the little electromagnetic unit on chip can be positioned at the zones of different on the chip.Spatially separate each other in these zones, such zone in owing to can not interfere with another zone for the electromagnetic force of electromagnetic units array generation.In the capable ripple design of magnetic force in the present invention, the magnetic field that different little electromagnetic units produce in the same area can temporarily overlappingly link to each other, and the magnetic field that array produced that is formed by the little electromagnetic unit of zones of different can temporarily overlappingly link to each other.
" manipulation " be meant and move or handle particulate, makes particulate do motion on one dimension, two dimension, the three-dimensional, and motion can occur on the chip piece, also can occur between the polylith chip.Manipulation to particulate in magnetic field comprises, but is not limited only to, and transports, adsorbs, catches, repulsion, enrichment, separation, differentiates, makes particle suspension or the like, also comprises the change to the particle movement direction.
" sample " is meant and anyly can therefrom separates or analyze the fluid of component.Sample can have various sources, can be from a biosome, and from a group biology of identical or different kind, also can be from environment, as take from a water body or soil, perhaps from food or industrial materials.Sample can be finished, also can be unprocessed.Sample can be a gas, liquid, and semisolid also can be solution or suspension.Sample can be a kind of extract, as takes from the liquid extract of soil or foodstuff samples, as throat or phallic extract, or as taking from the extract of faecal samples.Biological sample also can be organ, tissue, clone (comprising nutrient culture media and cell) or the like.In the present invention, Shou Xuan sample is a blood sample.
" blood sample " is meant at this and processed or unprocessed blood sample, such as, but be not limited only to, it can be the blood sample through centrifugal, filtration, extracting or other processing, can also be to have added anticoagulant, the blood sample of one or more reagent such as stabilizing agent.Blood sample can be an arbitrary volume, and the source is any, as animal or human's class.First-selected material is from the mankind.Blood can be directly extracts from biosome, also can be from the stock, and also can be from extracting on other object, for example clothes, furniture or the like.The volume of blood sample is not limit, and can decide according to concrete application greater than 5 liters less than 5 microlitres yet.
" leucocyte " is meant white blood cell, or a class can in animal blood, find neither the non-again hematoblastic hemopoietic cell of granulophilocyte.White blood cell can comprise lymphocyte, as bone-marrow-derived lymphocyte or T lymphocyte.White blood cell also can comprise phagocyte, as monocyte, and macrophage and include basocyte, eosinocyte, the granular white blood cells of neutrophil cell.White blood cell also comprises mast cell.
" red blood cell " is meant red blood cell.
" tumour cell " is meant the faster abnormal cell of a class cell proliferation growth fraction normal cell, it in addition can still continued growth after the stimulation that is subjected to causing new growth retardation.Compare with normal structure, oncocyte tends to partly or entirely lack structure organization and functional harmony, it both may be optimum also may be pernicious.
" malignant cell " is meant that a class has local invasion property and the destructive cell of growing and shifting.
" stem cell " is meant the cell that a class is broken up, and it can be taken turns or many wheel cellses differentiation causes growth by one, generates at least a noble cells.
" break up early stage cell " and be meant that a class is broken up but must will cause the cell of growth, it is taken turns by one or the differentiation of many wheel cellses causes growth, generates at least a noble cells.More representationally be, a stem cell responds to a special stimulation or a series of stimulation, take turns or early stage cell of differentiation of many wheel cellses differentiation generation by one, break up early stage cell by this then a special stimulation or a series of stimulation are responded, generate one or more noble cellss.
" pathogen " is meant any pathogen, as bacterium, virus, parasite or prion that can infection biological.Pathogen can cause being subjected to the biology of its infection disease million to occur or be in morbid state.Human pathogen is meant and can infects human pathogen.Human pathogen both can be that specificity is at the mankind, as the human pathogen of specificity; Can infect multiple biology again, as mixing property human pathogen.
" object " is meant any biosome, as animal or human's class.Animal can comprise any animal body, as wild animal, and domestic animal such as cat or dog, agricultural animal such as pig or ox, or animal such as horse are used in amusement.
" reaction tank " is a kind of structure of compositing chip, and it can the receiving fluids sample.
" port " is meant the opening of reaction tank body, and fluid sample can be by its turnover reaction tank.Port can be any size, but suitable be to allow sample by pipette, syringe, conduit or other add the shape and size that sample loading mode adds reaction tank.
" conduit " is in order to liquid is transferred to a kind of mode of reaction tank by container in this patent.The suitable port that is applied to reaction tank of conduit.Conduit can be made up of the material that any permission liquid passes through.Relatively Shi Yi conduit is a tubular structure, such as rubber tube, polyfluortetraethylene pipe or polyethylene pipe.Conduit can be an arbitrary dimension, but suitable be that internal diameter is between 10 microns to 5 millimeters.
" chip " is meant that can carry out one or many on it handles as physics chemistry, biochemistry, the solid matter of biological treatment.These processing can be to analyze, and comprise biochemical, cell and chemical analysis; Also separate, comprise making electricity consumption, magnetic, physics, (comprising biochemical) acting force or interaction of chemistry; Can be chemical reaction, enzyme reaction and association reaction also, comprise and catching.
" electromagnetic chip " is meant the chip that comprises an electromagnetic unit at least, for example only contains a little electromagnetic unit.Electromagnetic unit can be positioned at the surface of chip, also can be integrated or be partially integrated in the chip.For example, an electromagnetic unit can be produced on the surface of chip, also can be embedded in the chip, or be partially embedded in the chip.
" separation " is meant one or more components in the sample and the other spatially separated process of one or more components.Detachment process can be carried out like this, one or more interested entity molecules are located in certain or some position in the discrete device, at least other component in the sample segment is located in other position, or removes from the position at interested entity molecule place.Perhaps, one or more components in the sample can be positioned at some position, and one or more interested entity molecules are removed from this position, collect again.Also one or more components in the sample can be positioned at some position, again one or more other interested entity molecules be navigated to other position.Separation can realize by using physical force, chemical force, electric field force or magnetic field force or the like, can also use gravity field, fluid, dielectrophoresis force, row ripple dielectrophoresis force and electromagnetic force or the like to realize.
" catch " and be meant such class separate mode, this mode is trapped in one or more zones of chip one or more entity molecules, applies physical force again and catches.Catch and can realize by special capture probe, capture probe can be special, combine with interested entity molecule closely.These special capture probes can the reversible or irreversible solid dielectric that is connected on the chip on.
" analysis " is a test that component is carried out to sample or sample.The existence that analysis can detect certain component whether, the quantity of certain component or concentration, the composition of certain component, and the activity of certain component or the like.Analysis can be used with method of the present invention and device, reagent, comprises biochemical analysis, binding analysis, cell analysis and genetic analysis.
" reaction " is meant the chemistry or the biochemical component that can change one or more molecules or compound, perhaps changes the chemistry or the biochemical process of mutual relationship between one or more molecules and another or multiple molecule or compound.Reaction among the present invention can comprise by enzymatic, but is not limited only to, and degradation reaction arranged, synthetic reaction, modification reaction or association reaction.
" binding analysis " is a kind of analysis mode, it is by combining the predetermined substance molecule existence or the concentration that detects this predetermined substance molecule with specificity junction mixture, perhaps detect the possibility that combines of a predetermined substance molecule and another predetermined substance molecule, perhaps detect the binding affinity of a predetermined substance molecule and another predetermined substance molecule.The predetermined substance molecule can refer to an organic or inorganic molecule, and one by an organic and inorganic or organic-inorganic composite body, organelle, the molecule mixture that virus or cell are formed.Binding analysis can use detectable mark or can produce the signal generating system of detectable label in the presence of in conjunction with the predetermined substance molecule.The binding analysis of standard includes the dependence nucleic acid hybridization and detects special nucleotide sequence, relies on the combination of antibody to the predetermined substance molecule, and relies on the combination of part to acceptor.
" biochemical analysis " is meant a kind of existence of one or more components of sample, concentration or the active mode of analyzing.
" cell analysis " is meant a kind of analysis mode that detects cell processes, for example, but be not limited only to metabolic activities, catabolic activity, ion channel activity, intercellular signal conduction activity, receptor-mediated signaling activity, transcriptional activity, translation activity or secretion activity or the like.
" genetic analysis " is meant a kind of existence of certain hereditary unit or analysis mode of sequence of detecting.This hereditary unit can refer to any segment of DNA or RNA, comprises, but is not limited only to, and gene, repetitive sequence, transposable element, regulation and control unit, telomere, centriole or other have DNA or the RNA segment of not knowing function now as yet.The example of genetic analysis is as (but being not limited only to these examples), and nucleic acid hybridization technique comprises the nucleic acid sequencing reaction, can use one or more polymerases, for example the genetic analysis of PCR-based.Genetic analysis can be used one or more detectable marks, such as, but be not limited only to fluorescein, radioactive isotope or signal generating system.
" electrode " is a kind of structural unit of being made by high conductive material.Here so-called high conductive material is meant that the conductance of material is much larger than surrounding medium or material.High conductive material comprises metal, as gold, chromium, platinum aluminium or the like, also can be nonmetal, as graphite, conducting polymer.Electrode can be made different shape, as rectangle, circle, castellated or the like.Also can comprise doped semiconductor in the electrode material, as mixing phosphorus on the silicon chip.Can apply electric current on the electrode, it also is annular for example making the electric current that applies in the electrode of annular.
" groove " is the structural unit on the chip, has a lower surface and one or more side that extends out at a certain angle from the bottom surface.The parameter of side is very random, for example can have S shape or crooked or the side with a plurality of angles.The bottom surface of groove can be lower than, be higher than or and chip upper surface level.The material of side should be different with the material of the lower surface of chip.For example, the chip lower surface can be made of by the material that electric field force (comprising dielectrophoresis force, row ripple dielectrophoresis force and electromagnetic force) penetrates one deck, and the side of groove is made up of other material, as insulating material, can hinder penetrating of electric field force.The side of groove or the passage on the chip can be made by any suitable material, as silicon, glass, rubber, polymer, plastics, pottery, metal or the like.
" continuous flow " is meant in detachment process of the present invention, and fluid is pumped into or injects reaction tank by continuous.Like this, the component that is not trapped in the chip by selectivity in the sample will be taken out of reaction tank along with fluid.
" connector " is meant any any material that can link to each other with specificity with certain compatibility with entity molecule, and can handle by corresponding physical force.Connector can be, but be not limited only to cell, organelle, virus, particulate, condensate or complex, the perhaps condensate of molecule or complex.
" particulate " is meant Any shape among the present invention, and any composition has the material of any composite structure, can handle by magnetic force.Particle size can be from about 0.01 micron to about 1 centimetre.More suitably be, the size of the particulate in this method from about 0.1 micron to about several thousand microns.Particulate can be made by various suitable materials, and example comprises, but is not limited only to glass, pottery, polymkeric substance such as nylon, teflon (TEFLON TM), polystyrene, polyacrylamide, agarose, cellulose, glucosan or the like.The manufactured materials of the particulate that can be handled by magnetic force can be a magnetisable material, also can be can be by magneto motive material, for example iron, magnetite, ferromagnet and ferrimagnet material.
" coupling connection " is combination.For example, entity molecule can carry out special or non-specific combination with other particulate.In conjunction with being covalent bond, can be non-covalent bond also, be reversible combination also can the reversible combination of right and wrong.
" specific junction mixture " is meant in two such different moleculars, like this surface of molecule have specific morphosis can be specific and the space or the complementary combination of polar structure of another molecule.Specific junction mixture can be a kind of in two kinds of materials of specific bond in the immune family, as Ag-Ab, biotin-avidin, biotin-Streptavidin, ligand-receptor, double-strandednucleic acid, IgG-albumin A, DNA-DNA, DNA-RNA, RNA-RNA or the like.
" nucleic acid molecules " is meant polynucleotide.Nucleic acid molecules can be DNA, RNA or both combinations.Nucleic acid molecules also can comprise except ribose and ribodesose, form skeletal chain carbohydrate, can be the nucleic acid molecule outside DNA or the RNA.Nucleic acid molecules can be made up of the nucleotide base that exists naturally that exist naturally or non-, as xanthine, and the extension 2-aminoacyl purine of nucleoside base or analog etc.Nucleic acid molecules can be the peptide nucleic acid molecule.Nucleic acid molecules can be a random length, can be strand or two strands, or part strand and partially double stranded.
" mark that can be detected " is a kind of compound or molecule that can be detected, and they can produce output signal, the existing or method that can produce output signal that Future Development goes out as fluorescence, radioactivity, color, chemiluminescence or technical field.Output can be based on fluorescence, as by fluorescent marker, and Cy-3 for example, Cy-5, phycoerythrin, phycocyanin, allophycocyanin, fluorescein isothiocynate (FITC), rhodamine, or lanthanide series or the like, but be not only to be limited to above material; Also can be by fluorescin such as green fluorescent protein (GFP) and its variant, be based on the activity of enzyme reaction, such as, but be not limited only to β-nougat, beta-lactamase, horseradish peroxidase, alkaline phosphatase or luciferase or the like; Perhaps can based on radioactive isotope (as 33P, 3H, 14C, 35S, 125I, 32P or 131I etc.).Label also can be by other base group modification base, such as, modify pyrimidine in the C5 position or modify purine in the N7 position.Modification group can be diversified, for example halogen, ether or polyethers, alkyl, lipid or polyester, or common XR, and here, X is a linking group, R is a modification group.In the modification technique field, exist many kinds effectively may be applicable to the method (A Practical Approach, Eckstein, ed. (1991) and in PCT/US94/00193) of modified nucleic acid molecule, oligonucleotide molecules and analog thereof.
" signal generating system " can comprise one or more components, and one of them plants component is detectable bond.Signal generating system comprises that all are in order to produce or to strengthen the reagent that can survey signal (or act on mark signal to produce by certain method).Signal generating system provides a kind of mark that can use the external detection method to detect, and this method normally realizes by measurement absorbing light and radiative difference of wavelength.Signal generating system also can comprise chromogenic substrate and enzyme, and wherein chromogenic substrate absorbs the light of visible region or ultraviolet region or phosphorescence, fluorescence, thereby shows color under the effect of enzyme.Certainly, signal generating system also can use detectable label such as radioactive isotope so that detectable signal to be provided.
Signal generating system comprises at least a catalyzer (normally at least a enzyme) and at least a substrate, also can comprise two or more catalyzer and one group of substrate (such as comprising one group of enzyme, wherein a kind of substrate of enzyme is the product of another kind of enzyme).The operation of signal generating system is to produce detectable signal in predetermined site, promptly means in this predetermined site to have corresponding mark.
In order to obtain detectable signal, this just requires the signal that the mark at predetermined site produces is amplified.Like this, this mark generally is catalyzer, luminophor or radioactive isotope, preferably catalyzer.Catalyzer preferably can produce the enzyme or the coenzyme of multiple signal generation molecule from single marking.Enzyme or coenzyme can light absorbing products by producing, as dyestuff or be created in the product that irradiation can excite bright dipping down, as fluorescer to reach the purpose that desired signal is amplified.Perhaps, catalytic reaction also can directly produce exciting light, for example chemiluminescence.Many enzymes and coenzyme can be seen U.S. Patent No. 4,275 in order to this purpose, 149 and U.S.Pat.No.4, and 318,980 (in lists of references).In addition, many visible U.S. Patent No.s 4,160,645 of non-enzyme catalyzer (July 10,1979) that also can realize above-mentioned effect.
The product of above-mentioned enzyme reaction is dyestuff or fluorescer normally.In U.S. Patent No. 4,275, provided the explanation of a large amount of fluorescers in 149.
Other is used for herein term and has meaning consistent when using usually with this area, and explanation is all arranged in technical dictionary.
I. the electromagnetic chip that contains little electromagnetic unit
But of the present invention have a decline electromagnetic chip of electromagnetic unit of single-point gating, comprising: substrate; Be positioned at suprabasil a plurality of little electromagnetic unit; By to one or a plurality of little electromagnetic unit applies electric current, thereby produce method at the bottom of the magnetic field.Above-mentioned little electromagnetic unit can be a spatial structure, also can be planar structure.
Substrate
Substrate can be used any suitable chip production bottom material, for example, but is not limited only to silicon, glass, sintered glass, quartz, monox, plastics, pottery etc.Base material is preferably fine and close, but porous materials also can be used, especially the porosint of some operation among those suitable the present invention, these operations can be, but be not limited only to, association reaction, broken born of the same parents or detection association reaction etc., these action needs see through the base material transfer or transport material.
The size of substrate should meet the needs of micro fabrication, and micro-processing method commonly used comprises etching, sputter, photo etched mask or the like.The present invention that also should suit simultaneously of the size of substrate carries out the requirement of the method for little manipulation to particulate.For example, substrate can be a thin slice, about 1 millimeter of thickness, and length circle is in 5 millimeters to 1 centimetre, and width circle is in 10 millimeters to 5 centimetres.Substrate can be any suitable shape, and for example square, rectangle, ellipse, circle etc. also can be irregular set figures.First-selected shape is square, circle and suitable polygon among the present invention.
Substrate is the part with individual layer or multilayer chiop of multiple function.For example, can comprise multiple structure on the single layer of chips, to realize different functions.Vibration unit resemble the piezoelectric crystal can cause flowing of sample liquid, and the dielectrophoresis structure can be according to the different mobile entity molecules of entity molecule dielectrophoresis character.Certainly, these structures (for example vibrational structure and dielectrophoresis structure) liquid can be produced in the substrate of different layers.Among the present invention, these different basalises can in layer be connected together, form multifunction chip.These structures with difference in functionality can use suitable equipment independently to control.
Substrate be in the chip structure in order to hold sample, the reaction tank of fluid ingredient for example.Reaction tank can be a complete closed, though complete closed is not necessary.Reaction tank can by for example structure such as port or conduit permission sample or reagent carries out.Fluid or solid or carry out reaction tank by certain mode under the effect of certain power, for example by gravity or pump.Reaction tank also should have the structure of outlet simultaneously, and for example port or conduit leave to allow the material in the reaction tank.In one embodiment of the invention, reaction tank feeds material by the inlet of forming with port and guide-tube structure, derives material with the outlet that port and guide-tube structure are formed.
Little electromagnetic unit
Contain little electromagnetic unit on the electromagnetic chip among the present invention.This slightly electromagnetic unit comprise that one directly is produced on or is integrated in suprabasil magnetic core and can apply the method for electric current around this magnetic core.This magnetic core preferably has the core of magnetic or the core that can be magnetized.
The size of little electromagnetic unit
The size of little electromagnetic unit is relevant with the intensity in the magnetic field that needs to produce with the direction of little electromagnetic unit.Usually, the width of little electromagnetic unit and length circle are between 0.1 micron to 1 centimetre, and preferably the boundary is between 10 microns and 1 millimeter.That can disperse according to required magnetic field or be confined to a certain zone, select the size and dimension of little electromagnetic unit.Usually, opposite with electromagnetic unit in tape or these magnetic storage mediums of disk, disperse in the magnetic field that the little electromagnetic unit among the present invention produces, and has long relatively operating distance.
The position of little electromagnetic unit and direction
According to the concrete purposes of chip and the character in needed magnetic field, select concrete structure of little electromagnetic unit and position.Little electromagnetic unit can directly be produced on the surface of chip, or is partially embedded in the chip, or is embedded in the chip fully.Can modify layer of substance on the surface of substrate to protect little electromagnetic unit or to be used for fixing some material, for example entity molecule.
Each little electromagnetic unit can be made certain structure according to concrete needs, can be three-dimensional, and the part solid also can be the planar structure or the structure on part plane.The job operation that these structures are mentioned in can be according to the present invention is made.
Magnetic core
Any suitable material can be in order to make the magnetic core in little electromagnetic unit among the present invention.Usually adopt magnetisable material or the material that can be magnetized, for example ferromagnetic material or ferrimagnetism material.Magnetic core can use suitable method to make, and for example adopts the method for mentioning among the present invention.
Apply the method for electric current
Should comprise a circle or a multi-turn lead around the above-mentioned magnetic core.Lead around the magnetic core can be made different shape, can be circular, square, oval, triangle, scroll or the like.When applying electric current in individual pen around magnetic core or the multi-turn lead, will produce corresponding magnetic field according to electromagnet phenomenon.
The magnetic field that produces
The character in the magnetic field that is produced by the little electromagnetic unit of gating determines by multiple factor, for example the character of the shape of the direction of the magnetisable rate of the magnetic spectroscopy susceptibility of the ratio of the length of the size of magnetic core (length and width and height), magnetic core and sectional area, magnetic core, magnetic core, magnetic core, magnetic core and the electric current that applied.Among the present invention, disperse in desired magnetic field, rather than assemble, and this is opposite with the magnetic field properties that electromagnetic unit produced in tape or these magnetic storage mediums of disk.For example equal the United States Patent (USP) 5 submitted on March 16th, 1999 at Yamade et al., the ring that the structure of 833,760 described magnetic heads is made up of a circle magnetic material, the two ends of ring are very approaching, this special structure can produce the magnetic field of very strong gathering, rather than the magnetic field of dispersing.On the contrary, little electromagnetic unit of the present invention does not just have such structure, and in order to affact entity molecule or the magnetic particle that is distributed in zones of different, little electromagnetic unit of the present invention is related to becoming the structure that can produce divergent magnetic field.
Gating and modulation separately
Separately gating or close little electromagnetic unit among the present invention applies different electric currents, and the method for regulating the magnetic field that each little electromagnetic unit produces also belongs to category of the present invention.To the independent control of little electromagnetic unit is that lead, the power supply that is connected and control corresponding gating of drawing by each little electromagnetic unit and the switch of closing are realized.Switch can be a mechanical switch, also can be electronic switch, also can be the combination of above-mentioned switch.
Because each little electromagnetic unit is gating separately all, so at any one constantly, can have only a little electromagnetic unit is gating, produces single magnetic field, can a plurality of again little electromagnetic units of while be gatings also, the generation resultant field.Little electromagnetic unit of gating is meant certain electric current on this little electromagnetic unit, so that this little electromagnetic unit produces magnetic field in its peripheral region.Thereby attract or move the magnetic particle in its peripheral region or modified the molecule of magnetic particle for the magnetic field that make to produce has enough intensity, the amplitude and the phase place that are applied to the electric signal on little electromagnetic unit can be carried out suitable selection.As for not by little electromagnetic unit of gating, it may not produce magnetic field fully, can produce faint magnetic field yet, but the undercapacity in magnetic field is to attract or mobile magnetic particle or modified the molecule of magnetic particle.
Independent gating to little electromagnetic unit can be accomplished in several ways, and for example can all be connected to each end that comprises little electromagnetic unit of a conductor loop on the power supply, and the other end is connected with decision whether allow passing through of electric current separately with electric switch.In the ranks array that constitutes by little electromagnetic unit, a full line or a permutation all can be received together, thereby be realized gating one full line or a permutation.
Little electromagnetic unit among the present invention is gating separately, but is not all to need independent gating in all cases.Little electromagnetic unit can link together with suitable circuit, for example can be by in parallel or series connection.The little electromagnetic unit of on the chip each gating has separately brought great dirigibility for the application of chip.Only link together as the little electromagnetic unit on the fruit chip,, may make all unit all lose efficacy if one of them little electromagnetic unit breaks down in the mode of series connection; Only connect in parallel as the little electromagnetic unit on the fruit chip, though can address the above problem very inconvenience again in the time need carrying out the adjusting of independent gating and electric signal little electromagnetic unit.
The capable ripple of magnetic force of being expert at and using in the ripple magnetic force dielectrophoresis can be realized by little electromagnetic unit of gating separately of the present invention.For example, on chip, make the little electromagnetic unit of the wire that is parallel to each other of one group of planar structure, these unit the different time by gating, for example along certain direction gating successively, like this, magnetic field is along with the direction of gating is propagated, as wave propagation, similar with row ripple dielectrophoresis.Be expert in the ripple magnetic force dielectrophoresis, the capable ripple of magnetic force can form synchronous ripple by direct current, also can form continuous ripple by alternating current.Under the effect of the capable ripple of magnetic force, particulate can move according to mode and direction that the operator wishes on the chip.Xiaobo Wang, Weiping Yang, JunQuan Xu, the U.S. Patent application that Jing Chengand Lei Wu submitted on October 3rd, 2000 " Apparatus forSwitching and Manipulating Particles and Method of Use Thereof " (Patent Office's numbering: 09/678,263) proposed to use row ripple dielectrophoresis and particulate switch to carry out the method that particulate is handled on the chip.The present invention proposes and use capable ripple dielectrophoresis of magnetic force and particulate switch to carry out the method that particulate is handled on the chip.
When using direct current, little electromagnetic unit can be successively by gating with close.Be closed after little electromagnetic unit gating, can produce the demagnetization phenomenon, promptly magnetic field needs regular hour decay, for example sinusoidal decay.The electric current that applies during little electromagnetic unit gating should be between between the 0.1mA to 3A, better between between the 1mA to 300mA, preferably between between the 10mA to 30mA.
When using alternating current, can on each little electromagnetic unit, apply the electric signal of out of phase, for example continuous four little electromagnetic units respectively differ the phase place of 90 degree.The frequency of electric signal should be between between the 0.001Hz to 1GHz, be more preferably between between the 0.01Hz to 100MHz, or between between the 0.1Hz to 10MHz, or between between the 0.1Hz to 1MHz, or between between the 1Hz to 100KHz, or between between the 10Hz to 10KHz.
Array and size
Electromagnetic chip of the present invention comprises little electromagnetic unit structure, we recommend with this slightly electromagnetic unit arrange according to the arrangement mode of rule, and adjacent little electromagnetic unit is equidistant.
Little electromagnetic unit on the chip can be made according to same shape and size, also can make according to difformity and size.Generally speaking, the characteristic dimension of each little electromagnetic unit can be less than 1 micron, also can be greatly to 1 centimetre.Here, if little electromagnetic unit is circular, characteristic dimension is meant diameter of a circle; If little electromagnetic unit is foursquare, characteristic dimension is meant the foursquare length of side.This slightly electromagnetic unit can arrange according to the arrangement mode of rule, as the rectangular node shape, also can be arranged in irregular shape or randomize.
Functional layer
Electromagnetic chip of the present invention can comprise at least one one functional layer.On functional layer, can fix at least a entity molecule or aglucon.First-selected fixing entity molecule is nucleic acid molecules, antibody and acceptor.Among the present invention, functional layer can also be as the little electromagnetic unit or the protective seam of chip base material, in addition, can also provide may need in the experimental technique of the present invention chemical group.
Functional layer can be composed of any suitable material, and first-selected among the present invention is to comprise following a kind of material at least: hydrophilic unimolecular layer, the hydrophilic unimolecular layer that functional group is arranged, hydrophobic unimolecular layer, hydrophobic unimolecular layer, hydrophilic film that functional group is arranged, the hydrophilic film that functional group is arranged, hydrophilic gel, the hydrophilic gel that functional group is arranged, hydrophobic glue, hydrophobic glue, porosint that functional group is arranged, the porosint that functional group is arranged, non-porous material and the non-porous material of functional group is arranged.
Functional layer can be by adhering to, connect or being adsorbed on substrate upper base skim material, also can be that for example chemical modification forms on chip base by modification.Functional layer is by spraying in substrate, or base material is immersed in liquid or the liquid solid, makes these modify that materials solidify to form by modes such as cooling, gel polymerisation again.
Have on the functional layer a series of can be in order to the functional group of permanent entity molecule, first-selected functional group is, but is not limited only to acetaldehyde, charing diimine, succinimide ester, antibody, acceptor and lectin.These functional groups are modified material surface ready-made method is arranged.
Be fixed on the ingredient that entity molecule on the functional layer or aglucon also can be regarded functional layer as.First-selected fixing entity molecule or aglucon comprises, but be not limited only to nucleic acid molecules (for example strand or double-stranded DNA, RNA or their complex), set reagent (for example having in antibody or the antibody), acceptor, polypeptide, protein, carbohydrates, lipid, prokaryotic, eukaryotic, virus, parasite, prion, bacterial antibodies, agglutinin and acceptor or the like in conjunction with active segment.The functional layer that contains these entity molecules has the several different methods preparation.For example, had the appropriate functional group on the functional layer, the reagent that will contain entity molecule again is added on the functional layer, by functional group permanent entity molecule, also can pass through other method position determining entity molecule, for example uses the method for mask.
The making of little electromagnetic unit
When little electromagnetic unit of processing and fabricating passed to electric current on to base material, each unit can produce independently magnetic field.An example is exactly to be connected on the current source around a ring and by electronic switch with conductive material on ferromagnet or the iron core.This ring can be round, oval, square, triangle or other arbitrary shapes, and unique prerequisite is to want to form the electric current that flows around ferromagnet.If ring is a single turn, it should be closed or approaching closed.Ring also can be around ferromagnetic many circle circles.It also can be that another kind of situation is an independent stratum in each circle representative structure that these circles can be formed in same one deck of microstructure.Electric conductor can be the electrical lead that deposition forms---as through electroplating, and the metal construction that sputter or deposition form, electric conductor also can form by semiconductor layer is optionally mixed simultaneously.Little electromagnetic units array can be arranged in the common row-column configuration that has in the similar microelectronic component.Row and column can be vertical mutually, also can be arranged to other angles (as 80 degree).
Reaction tank
Reaction tank of the present invention is a kind of structure that can hold fluid.Reaction tank can be an arbitrary dimension, should hold the fluid of 1nl to 50ml usually, better should hold the fluid of 1 μ l to 10ml, and optimal cases is to hold the fluid of 10 μ l to 1ml.Generally speaking, reaction tank is one of structural unit on the chip.Reaction tank can be made by any suitable material, as silicon, glass, metal, pottery, polymkeric substance, plastics or the like, can be the quality stiff materials, also can be the material of quality softness.First-selected material is those not have the material of interference to dielectrophoresises motion of the entity molecule in the sample, for example those not with charged or the material that polar molecule combines, as the plastics or the polymer (as third rare acid amides, glass or the like) of silicon, some kind.
Reaction tank is one of ingredient of chip, and chip is a solid substrate, can carry out once thereon or separation repeatedly, analyzes, catches or the like process.The material of chip can be a kind of or several in metal, pottery, polymkeric substance, multipolymer, plastics, rubber, silicon, glass or the like.Chip can comprise several certain flexible materials that have.Chip can be made of porous materials, and also can be made by the material of densification.Be shaped on the chip or be integrated with such as the little processing structure of groove, passage, electrode unit or the like, can carry out reaction or processing physics, biophysics, biological, chemistry thereon.Chip is a thin slice.The chip surface area size is not done requirement, such as can from 1mm2 to 0.25m2.Preferably used chip surface area from 4mm2 to 25cm2 about.Chip can have difformity, shape such as rectangle, circle, ellipse or other irregular shape of rule.Chip surface can be flat, also can be uneven.The chip that has a non-planar surface can comprise from the teeth outwards by processing at chip surface or structures such as the groove that etching gets, reaction tank.
In the entity example, wrap electrode (electrode be produced on the chip or chip in) in the reaction tank, but this is not requirement of the present invention.Electrode on the chip can have arbitrary shape, for example rectangle, castellated, triangle, circle or the like.The arrangement mode of electrode also has many kinds, such as spiral fashion, parallel shape, interlaced shape, polynomial expression shape or the like.The making of the chip power utmost point can be used various little processing and methods of micro-mechanics common in this area, for example plating, metal sputtering, photochemical etching.The chip that includes electrode has, but be not limited only to, be produced on dielectrophoresis electrod-array (the Dielectrophoretic Manipulation of Particles by Wang et al. on the glass substrate, in IEEETransaction on Industry Applications, Vol.33, No.3, May/June, 1997, pages 660-669), have can single gate array little processing bioelectronics chip (Preparation andHybridization Analysis of DNA/RNA from E.coli on MicrofabricatedBioelectronic Chips by Cheng et al., Nature Biotechnology, Vol.16,1998, pages541-546) and capillary electrophoresis chip (Combination of Sample-Preconcentration andCapillary Electrophoresis On-Chip by Lichtenberg, et al., in Micro Total AnalysisSystems 2000 edited by A.van den Berg et al., pages 307-310).
Among the present invention or the reaction tank that uses in this method has one or more ports, or on the wall of reaction tank, has opening.Port can be any size, but suitable be to allow sample to add the shape and size of reaction tank by conduit.Conduit can be the tubular structure that any permission liquid passes through, such as rubber tube, polyfluortetraethylene pipe or polyethylene pipe.Port can provide an opening so that introduce sample by pipette or syringe for the reaction tank wall.
Conduit can be introduced sample from one or several port by pump (for example peristaltic pump or inculcate pump), syringe or gravity to reaction tank.One or more reagent, damping fluid, solution or the like.The sample solution that can modify the dielectric properties of one or more entity molecules in the sample among the present invention can add reaction tank simultaneously with sample, or adds reaction tank prior to, back in sample.Also can be before entering reaction tank, earlier sample and reagent, damping fluid or solution are mixed earlier, such mixed process can be carried out in the conduit of fluid being introduced reaction tank, also can carry out in one or a plurality of and sample cell that conduit links to each other.
II. the electromagnetic chip that has cavity
Electromagnetic chip with little electromagnetic units array of gating separately of the present invention is by forming with the lower part: substrate; In above-mentioned substrate, be arranged in the cavity of array; The ground floor lead, lead wherein is along the direction of the row of above-mentioned array; Second layer lead, lead wherein be along the direction and the ground floor lead mutually insulated of the row of above-mentioned array, and vertical mutually.Preferred option of the present invention is the mode that above-mentioned cavity is arranged in ranks, contains a magnetizable magnetic core in each cavity.
In the present invention, ground floor lead and second layer lead separate by the ground floor megohmite insulant.This ground floor megohmite insulant can be made by any suitable material, for example, but is not limited only to monox, silicon nitride, plastics, glass, pottery, photoresist and rubber.
Second layer megohmite insulant is deposited on the surface of second layer lead and magnetic core, and this second layer megohmite insulant can be made by any suitable material, can use and ground floor insulation course identical materials, also can use the material different with the ground floor insulation course.For example, but be not limited only to monox, silicon nitride, plastics, glass, pottery, photoresist and rubber.
Electromagnetic chip of the present invention can also comprise other conductor layer, and every lead wherein is the row or the direction of row in the array all, and and other conductor layer insulation.The lead of electromagnetic chip can be made by any suitable material, and first-selected material can be, but be not limited only to aluminium, gold, silver, tin, copper, platinum, palladium, graphite and semiconductor material.
Electromagnetic chip of the present invention can also comprise other functional layer.Functional layer can be composed of any suitable material, and first-selected among the present invention is to comprise following a kind of material at least: hydrophilic unimolecular layer, the hydrophilic unimolecular layer that functional group is arranged, hydrophobic unimolecular layer, hydrophobic unimolecular layer, hydrophilic film that functional group is arranged, the hydrophilic film that functional group is arranged, hydrophilic gel, the hydrophilic gel that functional group is arranged, hydrophobic glue, hydrophobic glue, porosint that functional group is arranged, the porosint that functional group is arranged, non-porous material and the non-porous material of functional group is arranged.First-selected functional group comprises acetaldehyde, charing diimine, succinimide ester, antibody, acceptor and agglutinin.
Electromagnetic chip of the present invention can comprise at least one functional layer and at least one fluid pool.Fluid pool can be introduced liquid makes it contact with array.Be manufactured with inlet/outlet or pipeline on the fluid pool, so that material, for example reagent flows into or leaves fluid pool.The design of the fluid pool of this inflow one deriving structure is particularly suitable for automation mechanized operation of the present invention.
First-selected electromagnetic chip
In figure and the instantiation is example with first-selected electromagnetic chip of the present invention all.Electromagnetic chip with first-selection of the little electromagnetic units array of gating (10) separately of the present invention is by forming with the lower part: substrate (16); In above-mentioned substrate (16), be arranged in the cavity (22) of array; Ground floor lead (30 '), lead have wherein centered on magnetic core at least and have reached more than 90 degree.Above-mentioned cavity is arranged in the mode of ranks, contains a magnetizable magnetic core (26) in each cavity.
Electromagnetic chip also comprises other conductor layer, and lead has wherein centered on magnetic core at least and reached more than 90 degree, this layer conductor layer and conductor layer No.1 mutually insulated, but connect by the line that runs through insulation course between the lead.The material of insulation course can be monox, silicon nitride, plastics, glass, pottery, photoresist and rubber.The material of conductor layer can be aluminium, gold, silver, tin, copper, platinum, palladium, graphite and semiconductor material.
Electromagnetic chip of the present invention can comprise the other insulation course that is deposited on array surface.This second layer megohmite insulant can be monox, silicon nitride, plastics, glass, pottery, photoresist and rubber.
Electromagnetic chip of the present invention can also comprise other functional layer.Functional layer can be composed of any suitable material, and first-selected among the present invention is to comprise following a kind of material at least: hydrophilic unimolecular layer, the hydrophilic unimolecular layer that functional group is arranged, hydrophobic unimolecular layer, hydrophobic unimolecular layer, hydrophilic film that functional group is arranged, the hydrophilic film that functional group is arranged, hydrophilic gel, the hydrophilic gel that functional group is arranged, hydrophobic glue, hydrophobic glue, porosint that functional group is arranged, the porosint that functional group is arranged, non-porous material and the non-porous material of functional group is arranged.First-selected functional group comprises acetaldehyde, charing diimine, succinimide ester, antibody, acceptor and agglutinin.
Electromagnetic chip of the present invention can comprise at least one functional layer and at least one fluid pool.Fluid pool can be introduced liquid makes it contact with array.Be manufactured with inlet/outlet or pipeline on the fluid pool, so that material, for example reagent flows into or leaves fluid pool.The design of the fluid pool of this inflow one deriving structure is particularly suitable for automation mechanized operation of the present invention.
III. the used method of detection reaction the present invention includes the method for guiding aglucon and target substance reaction.This method comprises the steps:
1) provide one to have a plurality of devices of little electromagnetic unit of gating separately, electromagnetic chip for example of the present invention;
2) on little electromagnetic unit functional layer is arranged, functional layer can directly contact with little electromagnetic unit;
3) modify the aglucon molecule and make that the aglucon molecule after modifying can be by the action of a magnetic field;
4) solution that will have an aglucon molecule of modification contacts with functional layer;
5) at least one little electromagnetic unit of gating optionally makes it produce magnetic field, locator qualification the aglucon molecule, make at least a portion aglucon molecule be fixed on the functional layer, thereby form by the fixing molecular Pareto diagram of aglucon;
6) modify target molecule and make that the target molecule after modifying can be by the action of a magnetic field;
7) will have target molecule solution and and the aglucon molecular action fixed; And
8) the little electromagnetic unit of gating optionally makes the aglucon molecular reaction of target molecule and predetermined fixed.
Chip
The first-selected electromagnetic chip that uses in the said method is the electromagnetic chip described in the present invention, and the chip of other kind also can use certainly.
Magnetic particle
The magnetic particle that uses among the present invention and the size of magnetisable material can be between micron order between the millimeter levels.Magnetic particle can be made up of various materials, can use various technology to make, as long as the magnetic field that biochip produces is enough to make that magnetic particle induces magnetic dipole.
The magnetic particle that can move under the electromagnetic force effect can be made up of magnetic material.First-selected material is a paramagnetic material, the magnetic dipole that it is induced by externally-applied magnetic field, and after externally-applied magnetic field removed, can reply was zero.Iron complexes is exactly a kind of suitable paramagnet.Among the present invention, the surface of magnetic particle can be wrapped by one or more compounds so that connect specific bond, in order to the direct or indirect interested entity molecule of catching.Magnetic particle among the present invention can be made Any shape, is made generally in sphere and elliposoidal.
The present invention can use several magnetic particles simultaneously.Different magnetic particles has different surface naturies, promptly they can with different entity molecule combinations, just can separate several different entity molecules simultaneously so by this method.The entity molecule of different surfaces character can be by at the different compound of the pan coating of magnetic particle, different bond in the reversible again or irreversible connection and being achieved.
Entity molecule to be handled can be by any feasible method and the surface combination of bond.For example, entity molecule can directly link to each other or link to each other by a linking arm with the surface of bond, preferably the linking arm that can cut.Entity molecule also can pass through covalency or non-covalent combination and bond surface combination.The connected mode on entity molecule and bond surface can be special, and is also can right and wrong special.Best, the connection between entity molecule and the bond can be cut, as the linking arm that can be cut off by chemistry, physics or enzyme reaction.
Linking arm (connecting key) is any predetermined substance molecule that entity molecule and bond can be coupled together.Such linking arm (connecting key) can comprise, but be not limited only to covalent bond between the amino acid or the connection of peptide bond, disulfide bond, thioether bond, interrupted thioether bond, the free radical (as amino and mercapto) or the like.The linking arm of other form comprises the linking arm that can be cut by acid, for example bismaleimideothoxy propane, these materials that can be cut under the slant acidity condition of acid labile-transferrin conjugates, adipic aciddihydrazide in cell; Also can be the glue crosslinking agent that can under ultraviolet light or visible light, be cut, constant regional CH1, CH2 in the IgG 1 and CH3 (Batra et al., Molecular Immunol., 30:379-386 ((1993)) for example.In some entity examples, can use multiple linking arm simultaneously, facilitate the use the advantage of various linking arms.The linking arm of other kind such as trityl linking arm come from trityl group, can discharge entity molecule (U.S.Patent No.5,612,474) under different Acidity of Aikalinity conditions.Also have some to connect with entity molecule (the Huston et al. that can vide infra, Proc.Natl.Acad.Sci.U.S.A., 85:5879-5883 (1988), Whitlow, et al., Protein Engineering, 6:989-995 (1993), Newton et al., Biochemistry, 35:545-553 (1996), Cumber et al., Bioconj.Chem., 3:397-401 (1992), Ladurner et al., J.Mol.Biol., 273:330-337 (1997) and in U.S.Patent.No.4,894,443).Usually the connected mode of selecting for use among the present invention is biotin-Streptavidin interaction, antigen-antibody interaction, aglucon-acceptor interaction or nucleic acid array complementation effect.The linking arm and the method that are used to connect entity molecule and particulate can be referring to U.S. Patent applications, and application number is 09/636,104 (submitting August 10 2000 date).
In some cases, when entity molecule-bond such as molecule-particle complex handled/be separated to preposition after, not next step reaction of disturbing molecule of the existence of particulate.Like this, just will not separate again with bond by molecule.But in some cases, such separation is again necessary.The feature of detachment process is by the character of entity molecule and bond, the surface appearance and the manipulation process decision of bond.Usually, the condition of the condition of separation and polymerization is just the opposite.For example, if the combination under high salt ionic concentration condition of entity molecule and bond, entity molecule can elute from bond under low salt ionic concentration condition so.Similar, if entity molecule was finished with being connected by specific linking arm of bond, the mode of Fen Liing is exactly to impose on the condition that this linking arm can specificity makes its fracture so.
Magnetizable material is meant when applying magnetic field can induce magnetic dipole, and when externally-applied magnetic field disappeared, the magnetic dipole that induces is vanishing thereupon also.In application, can use commercially available magnetizable material or magnetic particle.The size of majority of particles below micron (for example 50 nanometers to 0.5 micron) to tens microns.Particulate can have different structures and composition.A kind of structure of magnetic particle is at small bits of iron magnetic material outer wrapping one deck colloid, as polystyrene layer.Can also nano level ferrimagnet and colloid as, polystyrene mixes and forms.The surface of these two kinds of particulates all is a polystyrene, can be further with various types of molecular modifications in the above.
In the present invention, the solution that comprises magnetic particle also can comprise one or more bonds.Bond can be one or more protein, as antibody, antibody fragment, antigen, aglucon (for example, but being not limited only to receptor-ligand), phytohemagglutin phytolectin or the like.Bond also can be organic or inorganic molecule, for example the receptor-ligand of nickel, glutathione, biotin, Avidin, Streptavidin, non-albumen or aglucon analog or the like.Bond also can be a nucleic acid, as RNA, DNA or the nucleic acid of any non-natural existence.Bond can or irreversible combine reversible with magnetic particle.At solid phase surface coupling connection molecule, all be method common in this area as the method for nucleic acid and albumen.
Modify the aglucon molecule
The aglucon molecule can be the entity molecule of any kind of, for example, but is not limited only to, and biomolecule, chemical reagent, drug molecule, even can be ingredient, for example a cell of sample.Specifically, the aglucon molecule can be nucleic acid molecules, antibody/antigen or the like.
In order to use magnetic field that the aglucon molecule is positioned, moves, the aglucon molecule need be modified.The method of modifying has multiple, for example the aglucon molecule is connected with magnetisable material.The method that connects can adopt above-mentioned linking arm.Connection can be the form by covalent bond, also can be biological affine mode, biological example element-Avidin affinity interaction, agglutinin-heparin affinity interaction, acceptor-aglucon combination and antigen-antibodies effect.Best, this linking arm can cut, for example can be by light, heat, enzyme or the cutting of other chemical reaction.
In operating process of the present invention, the aglucon molecule can disintegrate down from magnetic particle, in fluid pool, the magnetic particle that disintegrates down can be under the effect of magnetic force cleared out by fluid and with the aglucon molecular separation.
Among the present invention, a kind of method of modifying the aglucon molecule is that the aglucon molecule is mixed with magnetic particle, and freezing aglucon molecule and the magnetic particle of making forms little solid magnetic particulate.In this case, can use the magnetic force sample spliter that this little solid magnetic particulate is arranged on the electromagnetic chip.
Modify target molecule
Target molecule can be the entity molecule of any kind of, for example, but is not limited only to, and biomolecule, chemical reagent, drug molecule, even can be ingredient, for example a cell of sample.Specifically, target molecule can be nucleic acid molecules, antibody/antigen or the like.
In order to use magnetic field that target molecule is positioned, moves, target molecule need be modified.The method of modifying has multiple, for example target molecule is connected with magnetisable material.The method that connects can adopt above-mentioned linking arm.Connection can be the form by covalent bond, also can be biological affine mode, biological example element-Avidin affinity interaction, agglutinin-heparin affinity interaction, acceptor-target combination and antigen-antibodies effect.Best, this linking arm can cut, for example can be by light, heat, enzyme or the cutting of other chemical reaction.
In operating process of the present invention, target molecule can disintegrate down from magnetic particle, and in fluid pool, the magnetic particle that disintegrates down can be cleared out by fluid under the effect of magnetic force and separate with target molecule.
Among the present invention, a kind of method of modifying target molecule is that target molecule is mixed with magnetic particle, and freezing target molecule and the magnetic particle of making forms little solid magnetic particulate.In this case, can use the magnetic force sample spliter that this little solid magnetic particulate is arranged on the electromagnetic chip.
Association reaction
In solution, intermolecular combination or reaction (for example, antigen and antibody; Special dna probe and its complementary sequence) be in the process of molecular diffusion, to realize by collision.The local concentration of the efficient of reaction and the speed and the molecule of participation reaction is relevant with the energy of collision.In the model of many biochips, a kind of molecule has been fixed on the surface of chip, and another molecule then is free in the solution of chip surface.When the molecule in the solution bumped with the molecule of fixing by passive diffusion, reaction took place.In the regular hour, often have only a fraction of molecule can spread also and bump with fixing molecule.Like this, sluggish, inefficiency has had a strong impact on speed, efficient and the sensitivity of biochemical reaction on these biochips.And in the electromagnetic chip of the present invention, the molecule in the solution under the effect of magnetic force initiatively be fixed on the chip surface molecule and contact, can improve speed, efficient and the sensitivity of reaction greatly.
With a magnetic particle of being made by the superparamagnetism material is example, and when particulate is in the magnetic field B, particulate itself can be brought out generation magnetic dipole Φ =V p(∏ p-∏ m) H mV wherein pRepresent the volume of particulate, χ pAnd χ mIt is respectively the polarizability of particulate and surrounding medium.Φ mBe the magnetic spectroscopy susceptibility of medium, H mRepresent magnetic field intensity.Be shown below the magnetic force F that particulate is subjected to MagneticThe polarizability and the magnetic field gradient that depend on magnetic dipole:
F Nagnetic=0.5V p(χ p-χ m) H mΛ B m, wherein " X " and " Λ " represents dot product and gradient computing respectively.Whether particulate is subjected to the difference that magnetic force depends on the polarizability of particulate and its surrounding medium.Usually particle suspension (polarizability is close to zero) in the non-magnetic media of liquid state like this in order to produce enough magnetic force, just needs to use magnetic particle (polarizability substantially exceeds zero).Magnetic particle speed ν under magnetic force and viscous resistance effect during balance ParticleFor ν particle = F magnetic 6 π η m R is the magnetic particle diameter in the formula, η mCoefficient of viscosity for surrounding medium.Like this, in order to obtain big as far as possible magnetic field steering force, should consider following factor: the magnetic susceptibility of (1) particulate should be big as far as possible; (2) magnetic field intensity should be big as far as possible; (3) magnetic field intensity gradient should be big as far as possible.
Detect
The combination of aglucon molecule and target molecule can detect by several different methods.Wherein a kind of method is to use detectable label or uses detection system.Detectable label or detection system can produce visible signal or the visible signal of optics.In order to detect the combination of aglucon molecule and target molecule, can or connect the part that goes up detection system with the detectable mark of mark on aglucon molecule or the target molecule.For example, can feed the antibody of tape label in fluid pool, these antibody can be by specific bond fixing on the chip, and as the antigen combination, unconjugated reagent can be by the fluid flush away.According to the type of employed mark, can use suitable method and instrument, for example by optical fiber or CCD Equipment Inspection optical signalling, use magnetic resonance head (MR Head) can detect magnetic signal.
IV. the method for operating magnetic particulate
The present invention includes the operating magnetic particulate or the method for magnetizable particles.The step of method of operating is: provide one to have a plurality of electromagnetic chips of little electromagnetic unit of gating separately; Magnetic particle or magnetizable particles are added to the surface of chip electromagnetic chip; One of gating or a plurality of little electromagnetic unit as required.The electric signal that adjusting applies with the Distribution of Magnetic Field on change electromagnetic chip surface, thereby changes the magnetic field force that is subjected on magnetic particle or the magnetizable particles.Like this, magnetic particle or magnetizable particles can be moved to the zone of setting or leave from the zone of setting.
Magnetic particle or magnetizable particles comprise at least a entity molecule, comprise the ingredient of sample, for example cell (haemocyte, malignant cell, tumour cell etc.).Entity molecule can also be nucleic acid molecules, special binding reagents (for example antibody and acceptor).First-selected entity molecule is that the ingredient of nucleic acid molecules, DNA, RNA, acceptor, polypeptide, protein, carbohydrates, lipid, prokaryotic, eukaryotic, virus, parasite, prion, bacterial antibodies, agglutinin, acceptor or sample comprises cell, for example haemocyte, malignant cell, tumour cell etc.
Entity molecule is connected with magnetic particle or magnetizable particles.Being connected of magnetic particle or magnetizable particles and entity molecule can be by direct mode, also can be by non-direct mode.Non-direct mode can be passed through accomplished in many ways, for example passes through the method for gathering, or by special bond (antibody, acceptor or the like).Directly connection also can pass through accomplished in many ways, for example use chemical linking arm with magnetic particle or magnetizable particles with the entity molecule connection or directly be coupled at.
Zone specific on the chip can be shifted to or leave to the particulate that is connected with entity molecule under the effect of magnetic force.Be connected with the surface that is fixed on chip that the particulate of entity molecule can be reversible under the effect of magnetic force, can clean chip like this,, just can feed fluid and clean as fluid pool is arranged on the fruit chip.In addition, the functional group of chip surface, the functional group on the functional layer for example also can be reversible or irreversiblely combine with entity molecule, particulate or entity molecule-particle complex, thereby make the cleaning to chip become possibility.Like this, just can realize separation and location to entity molecule.
In case entity molecule is by reversible or irreversible separation, location or after fixing, several different methods can be used for the detection to it.For example, can be detected for an entity molecule of having located or particulate by detectable mark or detection system on it.First-selected detectable mark comprises fluorescence labeling, visible light mark, isotope labeling or the like.Detectable mark can just be connected on the entity molecule before separating, also can be at after separating, and by secondary reagent, for example labelled antibody is connected on the entity molecule.Also can have mark on the particulate, this mark can be the intrinsic character of particulate itself, and for example color etc. also can be the mark of modifying up afterwards, for example fluorescence labeling etc.According to the type of employed mark, can use suitable method and instrument, for example by optical fiber or CCD Equipment Inspection optical signalling, use magnetic resonance head (MR Head) can detect magnetic signal.The method that detects can be quantitative, semiquantitative or qualitatively.
For the entity molecule by magnetic separation is detected, should have detectable mark at least a entity molecule in the sample.For example, biological sample and sample solution of the present invention being mixed when carrying out magnetic separation, can use a kind of cell or the antibody of component in the specific identification sample that certain cell or component are carried out mark.Can have detectable mark on the antibody, for example fluorescence molecule is (as rhodamine, fluorescein, red, the phycoerythrin of Texas, phycocyanin, green fluorescent protein, red fluorescent protein, blue fluorescent protein, yellow fluorescence protein or the like.Cell also can use and have the not different antibodies mark of isolabeling.Like this, having the residing position of fluorescently-labeled cell can be detected, and uses sample solution sample component to be carried out the effect of magnetic separation thereby can judge.Except extracellular sample component, can utilize antibody to carry out mark as organelle, virus, protein, compound and nucleic acid or the like, thereby detect their magnetic separation.
In addition, also having a kind of detection mode is at after separating, detects the existence of protein, nucleic acid and other compound by binding analysis.For example, after sample solution of the present invention and sample mix, carry out cell separation again, the effect of separation can or detect with the antibody of the expressed protein-specific combination of given cell and can obtain with the probe nucleic acid sequence of distinguished sequence combination in definite cell (as the bacterium of special kind) or the like method by detecting.Can or analyze (analyzing) by enzymatic testing process (as PCR) to representing the special cell type or the detection of the nucleotide sequence of specific cell component and protein as P450.Can pair cell magnetic separation process monitor by introduce detectable mark (as dyestuff) to cell interior.For example, can introduce BCECF-AM (Molecular Probes in the cell, Eugene, OR), this fluorescein probe can be absorbed by living cells, after magnetic separation, (Gascoyne et al.IEEE Transcactions 33:670-678 (1997)) can be determined in the position of this fluorescein molecule.Chip in order to isolated cell is proved to be and can observes with microscope, separated like this entity molecule can be washed out reaction tank, analyzes by micro measurement, Flow Cytometry, cell growth analysis or the like method (but be not limited only to these several) again.
The entity molecule of having located can also be further processed.For example, can entity molecule be disintegrated down from particulate by suitable method.For example, can cut down by suitable method, for example cut by chemical reagent, enzyme, pH, salt or light in order to the linking arm that connects particulate and entity molecule.
Entity molecule to be handled can be by any feasible method and the surface combination of bond.For example, entity molecule can directly link to each other or link to each other by a linking arm with the surface of bond, preferably the linking arm that can cut.Entity molecule also can pass through covalency or non-covalent combination and bond surface combination.The connected mode on entity molecule and bond surface can be special, and is also can right and wrong special.Best, the connection between entity molecule and the bond can be cut, as the linking arm that can be cut off by chemistry, physics or enzyme reaction.
Linking arm (connecting key) is any predetermined substance molecule that entity molecule and bond can be coupled together.Such linking arm (connecting key) can comprise, but be not limited only to covalent bond between the amino acid or the connection of peptide bond, disulfide bond, thioether bond, interrupted thioether bond, the free radical (as amino and mercapto) or the like.The linking arm of other form comprises the linking arm that can be cut by acid, for example bismaleimideothoxy propane, these materials that can be cut under the slant acidity condition of acid labile-transferrin conjugates, adipic aciddihydrazide in cell; Also can be the glue crosslinking agent that can under ultraviolet light or visible light, be cut, constant regional CH1, CH2 in the IgG 1 and CH3 (Batra et al., Molecular Immunol., 30:379-386 ((1993)) for example.In some entity examples, can use multiple linking arm simultaneously, facilitate the use the advantage of various linking arms.The linking arm of other kind such as trityl linking arm come from trityl group, can discharge entity molecule (U.S.Patent No.5,612,474) under different Acidity of Aikalinity conditions.Also have some to connect with entity molecule (the Huston et al. that can vide infra, Proc.Natl.Acad.Sci.U.S.A., 85:5879-5883 (1988), Whitlow, et al., Protein Engineering, 6:989-995 (1993), Newton et al., Biochemistry, 35:545-553 (1996), Cumber et al., Bioconj.Chem., 3:397-401 (1992), Ladurner et al., J.Mol.Biol., 273:330-337 (1997) and in U.S.Patent.No.4,894,443).Usually the connected mode of selecting for use among the present invention is biotin-Streptavidin interaction, antigen-antibody interaction, aglucon-acceptor interaction or nucleic acid array complementation effect.The linking arm and the method that are used to connect entity molecule and particulate can be referring to U.S. Patent applications, and application number is 09/636,104 (submitting August 10 2000 date).
In some cases, when entity molecule-bond such as molecule-particle complex handled/be separated to preposition after, not next step reaction of disturbing molecule of the existence of particulate.Like this, just will not separate again with bond by molecule.But in some cases, such separation is again necessary.The feature of detachment process is by the character of entity molecule and bond, the surface appearance and the manipulation process decision of bond.Usually, the condition of the condition of separation and polymerization is just the opposite.For example, if the combination under high salt ionic concentration condition of entity molecule and bond, entity molecule can elute from bond under low salt ionic concentration condition so.Similar, if entity molecule was finished with being connected by specific linking arm of bond, the mode of Fen Liing is exactly to impose on the condition that this linking arm can specificity makes its fracture so.
When entity molecule is cell, can carry out certain processing by pair cell, for example make cell break born of the same parents by cracking or by the osmotic pressure that changes cell environment of living in.By these modes, can so that cell or intracellular entity molecule discharges, for example nucleic acid molecules, protein, polypeptide, acceptor, film fragment and organelle or the like.Like this, intracellular interested entity molecule just can obtain separating from sample.Entity molecule can be collected and be further processed in the cell.Easier realization above-mentioned purpose when chip adopts the pattern of fluid line.Intracellular material can detect by suitable method, for example can adopt PCR or the detection of the method for hybridization for nucleic acid molecules, for antigen, can adopt the method for immunoassay to detect.
Can adopt different marks to detect for different entity molecules.For example, can detect a kind of entity molecule, use fluorescein to detect another entity molecule with the Texas red marker.Can further detect on the chip piece or on the polylith chip at the entity molecule that separates on the chip.On chip, the specific site that can catch entity molecule is arranged.These entity molecules comprise special bond for example antibody, aglucon, acceptor or nucleic acid molecules, by some mode, and for example effect of functional layer, these entity molecules are fixed on these specific sites by reversible or irreversible mode.These style of a verse, poem, etc. molecules can be positioned on the chip by several different methods, for example by method of printing, comprise the contact point sample and use the ink-jetting style point sample.These different entity molecules can adopt identical or different marks to detect.The result of reaction detection can show sample information.By the result of comparative sample and contrast, can determine the absolute or relative concentration of component in the sample.This detection method is had many uses, for example be used for disease condition diagnosis, be used for the number change situation of the multiple entity molecules of research such as monitoring of environmental detection, medicine toxicology, medicine genetics, genomics.
The present invention has used electromagnetic running wave, also can be called capable ripple magnetophoresis (traveling wavemagentophoresis).Row ripple magnetophoresis is meant magnetic particle or the motion of magnetizable particles under the capable ripple effect of magnetic force.Capable ripple magnetophoresis like this can the application of the invention method and device realize.Magnetophoresis can be synchronous, also can be continuous.In synchronous magnetophoresis, direct current can be applied on the electromagnetic unit according to a certain direction successively, makes electromagnetic unit produce magnetic field successively, forms the capable ripple of magnetic force, and magnetic particle or magnetizable particles just can be moved along this direction like this.In continuous magnetophoresis, the ac signal that is applied on the electromagnetic unit differs certain phase place each other, and for example per 4 adjacent electromagnetic units differ the phase places of 90 degree.So also can form the capable ripple of magnetic force.
Device shown in Figure 24 B, the electromagnetic unit (190) that is arranged in capable ripple magnetophoresis device (170) show gating separately.On the chip to the circuit connecting mode of electromagnetic unit make electromagnetic unit can be separately by gating with close, the mode of gating successively for example: at first, first electromagnetic unit is by gating; Subsequently, second electromagnetic unit (the immediate electromagnetic unit parallel with first electromagnetic unit) is by gating, and first electromagnetic unit is closed simultaneously; Be that second electromagnetic unit closed then successively, the 3rd electromagnetic unit gating simultaneously ... like this, first, second, the 3rd ... just occur magnetic field on the electromagnetic unit successively, along with certain direction is propagated, form the capable ripple of magnetic force as magnetic field.These electromagnetic units have formed a capable wave structure of magnetic force.Under the effect of the capable ripple of magnetic force, the magnetic particle or the magnetizable particles that have or do not have entity molecule just can move along the direction in magnetic field.
A capable ripple magnetophoresis device comprises the electromagnetic unit of some.As long as can be suitable for row ripple magnetophoresis, these electromagnetic units can be size, shape and intensity arbitrarily.Generally speaking, row ripple magnetophoresis device comprises 2 to 1000 electromagnetic units, is more preferably to comprise 5 to 500 electromagnetic units, preferably comprises 10 to 100 electromagnetic units.Electromagnetic unit can be made any suitable structure and size, also will have the magnetic core of some so that produce enough magnetic field.The factor that need consider when making electromagnetic unit comprises the character of the intensity of quantity, the unit of size, the unit of unit, the particulate that need be moved and the character of the electric signal that need apply.The present invention can be in order to instruct design, to make and test multiple such structure, to determine the structure that can be used for row ripple magnetophoresis.
Among the present invention, different bonds can be fixed on simultaneously that (for example on functional layer) has the different loci of capable ripple magnetophoresis structural region on the chip.The particulate that has an entity molecule can pass through chip upward traveling wave magnetophoresis structural region by the effect of row ripple magnetophoresis, and like this, the be expert at different bond in ripple magnetophoresis zone of predetermined fixed can be caught each self-corresponding bond.In the ripple magnetophoresis process of being expert at or after row ripple magnetophoresis finished, the entity molecule of catching can adopt identical or different certification marks or detection system detects.According to employed mark, can select for use suitable detection method to detect.First-selected mark mode is a fluorescence labeling, and first-selected detection mode is to use optical fiber and CCD equipment, and concrete method is to use optical fiber structure to collect fluorescence signal, is transferred to CCD and partly measures and handle.
Example example 1: electromagnetic chip with little electromagnetic unit of vertical structure
It below is an example with electromagnetic chip of the little electromagnetic unit of vertical direction of the present invention.
Fig. 1 is the sketch of electromagnetic chip 10 among the present invention.Chip 10 comprises a plurality of little electromagnetic units 11, and these unit can be produced on the substrate 16 of the fine and close and non-dense material of silicon, glass, monox, plastics, pottery or other.Electromagnetic unit 11 on the chip 10 is arranged in 3 * 3 array.Electromagnetic unit 11 can induce magnetic field when applying electric current 15 17, and can carry out gating by multiple means.Fig. 1 is near the synoptic diagram that produces magnetic field behind 6 gatings in 9 little electromagnetic units each unit.The direction in attention magnetic field depends on the direction of electric current circulation.
In Fig. 1, electromagnetic unit 11 can be by go in ring electrical lead (annulus 15 as shown in the figure) and these electrical leads around a center 19 of insulating with the galvanic circle constituted.Ring can have multiple geometric configuration, as annulus, helix, square and square spiral shape.As being familiar with known to micro etch and the micro-machined personnel, these have different in width and thickness lead can preparation (can be referring to Rai-Choudhury P chief editor's Handbook ofMicrolithography on silica-based with different photoetching and processing scheme, Micromaching and Microfabrication, Volume 2:Micromachingand microfabrication.SPIE Optical Engineering Press, Bellingham, Washington, USA, 1997).These schemes comprise many basic steps, as making mask blank, plated metal, deposition insulator, apply photoresist, forming pattern, metal or insulation course moulding by mask and developer on photoresist.Lead can be metal material such as aluminium, gold, silver, tin, copper, platinum, palladium or nonmetallic materials carbon, and semiconductor material is as mixing phosphorus silicon, or other any materials that can conduction.In order to conduct the electric current up to the hundreds of milliampere, lead must have the cross-sectional area of several thousand square microns.The thickness of lead and width can change between 0.1～500 micron and 1～500 micron respectively.To each electromagnetic unit, lead can be single-turn or multi-turn.As be multi-turn, might need to adopt the little processing scheme of multilayer to prepare these unit.
In a kind of chip design scheme of the present invention, the gating of little electromagnetic unit links to each other the galvanic circle by electric switch and realizes with current source.Current switching situation in the galvanic circle can be applied to signal controlling on the electric switch by change, thereby makes electromagnetic unit or connection or shutoff.In another kind of chip design scheme, the gating of little electromagnetic unit can be realized by the current switching that mechanical switch is controlled the galvanic circle.Electromagnetic unit all is connected with switch in above-mentioned these two kinds of forms, just can realize the multiple combination of electromagnetic unit on/off strobe state by " opening " or " pass " of gauge tap.
In order to increase the magnetic field intensity that electric current is responded in conductive coil, can adopt the magnetic core of Ferrite Material or ferromagnetic material.In this case, each electromagnetic unit has all comprised the single-turn or multi-turn lead of on-chip magnetic core, wounded core, has applied the device of electric current from the current source guiding line.Therefore, the center 19 of electromagnetic unit 11 can be made by Ferrite Material or ferromagnetic material in Fig. 1, and makes itself and current return 15 carry out electrical isolation.Can be used on substrate deposited iron ferrite or ferromagnetic material at the known the whole bag of tricks of this field people and (see the article of Ahn and Allen for example, " A new toroidal-meander type integrated inductor with a multilevel meander magnetic core ", IEEETransations on Magnetics 30:73-79,1994).
The synoptic diagram of Fig. 2 for magnetic particle 21 is guided to an electromagnetic unit of working 11.When applying electric current 15, can produce induced field near this unit
Figure A0113635000471
17, this magnetic field produces magneticaction to particulate.As shown in Equation 3, magnetic force greatly depends on magnetic field
Figure A0113635000472
(and field intensity
Figure A0113635000473
) distribution.By just may command and change DISTRIBUTION OF MAGNETIC FIELD of gating electromagnetic unit selectively.For example, four the adjacent electromagnetic units of electric current synchronization motivationtheory by a certain suitable direction just can produce a quadrapole magnetic field.The current amplitude and the direction that are applied on little electromagnetic unit by change can further change DISTRIBUTION OF MAGNETIC FIELD.The change in magnetic field and then influence influence position, speed and other kinetic parameters of particulate to the magnetic force size that magnetic particle produces.For example, by increasing magnetic field intensity and magnetic force the speed of particulate is increased.
Electromagnetic biological chip shown in Figure 3 except the functional layer 42 that is positioned at chip surface other all the chip with shown in Figure 1 is identical.This functional layer is used for fixing the aglucon molecule, and it can be the combination of hydrophilic unimolecular layer or hydrophobic unimolecular layer, hydrophilic or hydrophobic film, hydrophilic or organophilic gel layer, polymeric layer, non-densification or dense material and/or these materials.Molecular monolayer is meant unimolecular layer (as the Langmuir-Blodgett film).Be the fixed nucleic acid aglucon, can use bond material used in Southern blotting and Northern blotting such as nitrocellulose or nylon.Albumen and polypeptide can come immobilization (for example hydrophobic) in addition by various physics or chemical means.For example, specific acceptor such as antibody or lectin can be added on the functional layer 42 in order to solidify albumen or polypeptide aglucon molecule.The aglucon of Gu Huaing reaches reaction and the analysis that will carry out on biochip as required, different molecules can be added on the functional layer 42, to reach the purpose of immobilization aglucon.These are called functional group for the molecule that fixing aglucon molecule is incorporated on the functional layer 42.Functional group can be (but being not limited to) acetaldehyde, carbodiimide, succinimide ester, antibody, acceptor and lectin.These functional groups also comprise by chip surface being carried out chemical group or the molecular locus that chemical modification forms.The using method of electromagnetic biological chip 10 shown in Figure 3 will specifically describe in the back.
Fig. 4 is the simplified diagram of little electromagnetic biological chip 10 of providing according to a kind of design proposal of the present invention noted earlier.Carry out conductivity by conductor 14 tie points 12 with electromagnetic units array.Fig. 5 has provided the cross sectional representation of detailed single electromagnetic unit.Though can produce similar little electromagnetic chip on the substrate in a lot, the substrate of anticipating shown in the illustrated scheme is the silicon chip 16 of surface finish.Below we describe the manufacturing process of electromagnetic biological chip 10 among Fig. 4 in detail.These processes only supply the usefulness of signal.Know micro-machined people and can revise these steps or process at an easy rate, improve and the part job operation with biochip of same structure shown in Figure 4.Conductive region is by surface diffusion (doping) the phosphorus resistance that to produce a doping content be 2-10 Ω/.Silicon dioxide insulating layer has the thickness of 1000-8000 , and produces by following described thermolysis process.
According to the size of little electromagnetic units array chip and the requirement of array density, form parallel wire 18 by mixing phosphorus photoetching on substrate 16.The superficial density of regulating phosphorous diffusion makes the square resistance of lead 18 be less than or equal to 10 Ω/.Because lead 18 forms in substrate 16, they do not have to rise and fall and can not exceed the surface of substrate 16 yet.
After forming ground floor lead 18, chip is placed on (as 1000 ℃) in the high temperature oven, be the insulating layer of silicon oxide of 2000-4000 at superficial growth one bed thickness of substrate 16.Cover ground floor lead 18 so on substrate 16, form ground floor insulation course 20.
Use the method for photoetching, the zone of appointment etches the aperture that is used to electroplate between the ground floor lead.For example, be that the plating pitting array 22 of 10 μ m is to form by add KOH solution (30%w/w) corrosion on silicon chip 16 surfaces deeply.Each xsect of electroplating pitting 22 is trapezoidal, and less parallel edges is near the lower surface of substrate 16.Another bed thickness is that the silicon dioxide 24 of 5000 is deposited on the surface of electroplating pitting 22, and will electroplate the layer of silicon dioxide removal of pitting 22 bottoms by photoetching.
In aperture 22, fill ferromagnetic material to form magnetic core.Earlier substrate 16 is placed on NiSO 4(200-400g/l) in the solution, under the situation of logical nitrogen, be warmed up to 400-600 ℃ of heating 30 minutes, so as to form in the bottom of electroplating hole 22 one thick be the nickel Seed Layer of 1 μ m.
Magnetic core 26 in the pitting 22 can form by electroplating by following step and condition: (1) 20-40 ℃ is placed down in Fe/FeCl 2In the solution (ratio 200:500g/l); (2) under 30-60 ℃ at FeNi/NiSO 4In the solution (200:400g/l); (3) under 30-60 ℃ at FeCl 2In the solution (10-60g/).Like this, form the array of magnetic core 26 on substrate 16, wherein the surface of the surface ratio ground floor silicon dioxide insulating layer 20 of magnetic core 26 wants high.Electroplate the different magnetic core 26 of acquisition constituent structure according to other conditions and step.For example, for obtaining not alloy of nickel (81%)-iron (19%) glass, electroplating solution can comprise following component: N iSO 46H 2O (200g/l), FeSO 47H 2O (8g/l), NiCl 26H 2O (5g/l), H 3BO 3(25g/l) and Saccarin (3g/l).～5mA/cm 2The current density condition under can obtain～electroplating velocity of 0.3 μ m/min.The details of other relevant plating conditions can (for example find from many lists of references, Romankiwand O ' Sullivan, " Plating techniques " in Handbook of Microlithography, Micromaching and Microfabrication, Volume 2:Micromaching andMicrofabrication, Editor:Rai-Choudhury P., SPIE Optical Engineering Press, Bellingham, Washington, USA, 1997).
After forming array of cores 26, a layer thickness is about the Si of 5000 3N 4Insulation course 28 is being deposited under 200-300 ℃ the temperature on the magnetic core 26 and first insulation course 20.Next be that the conduction aluminium lamination that thickness is about 1.2 μ m is splashed to Si 3N 4On the surface of insulation course 28.Between magnetic core 26, form the second layer lead 30 vertical by aluminium being carried out photoetching and wet etching with ground floor lead 18.So just, formed a little electromagnetic units array that is constituted by array of cores and bidimensional traverse net.The surface of aluminum conductor 30 may maintain an equal level or the summary height than the surface of magnetic core 26.
At last, second thickness is about the Si of 4000 3N 4Insulation course 32 is deposited over the surface of aluminum conductor 30 under 300 ℃.Then, the insulating material of ground floor lead 18 and second conductor layer, 30 ends is removed, made the end of lead link on the bonding wire point 12, thereby the lead on the chip is linked to each other with external circuit by conductor 14 by dry etching.
The conductive channel 18 and 30 of little electromagnetic units array is by DC power supply.Each unit in little electromagnetic units array can be controlled by giving different conductive paths 18 and 30 energisings selectively.As shown in Figure 5, by selecting the sense of current in magnetic core 26 passage 18 and 30 on every side to make around magnetic core 26 formation ring currents, can near this unit, produce a magnetic field, that is to say, in order to magnetize arbitrary some magnetic cores that list, must be selected and allow and pass to the opposite electric current of direction in these two passages at the conductive channel 18 of this row left and right side.Certainly, this electric current will magnetize magnetic cores all in these row to a certain extent.But, this to list arbitrary predetermined unit also be simultaneously a unit in certain delegation.When flowing through in the conductive path 30 of electric current in these row both sides, all component units on this row all can have magnetization to a certain degree.But for selected cell, will in the conductive path around it, there be electric current to flow and forms a loop as shown in Figure 5.The magnetic field intensity that produces in the selected cell place will be those not twices of selected unit like this.
When needs strengthen the magnetic field intensity of selected unit, can make many " circle " conductive channel (as miniature coils) with increase magnetic core around electric current.By can on insulation course 32, making one or more layers two-dimentional conductive channel net again with making passage 18 and 30 similar methods, each stratum reticulare comprises the two-layer conductive channel to 18 mutually insulateds similar with 30, and their position and passage 18,30 difference correspondences.
The magnetic field intensity of selected unit can also be improved by little coil that Micrometer-Nanometer Processing Technology is made wounded core.Under the certain situation of electric current, be directly proportional with the number of turn of miniature coils by the magnetic force that magnetic core produced.For the personnel that are engaged in microfabrication, they will be appreciated that has many Micrometer-Nanometer Processing Technologies all can be used to make miniature coils in the existing method.Following method is that the example of this patent is described, but method for making has more than and is confined to that this is a kind of.The method for making of miniature coils and the method for making of above-mentioned conductive channel are similar, also be or adopt doped conductive layer or adopt metal conducting layer (as aluminium) its difference is between the conductive layer it is that direction perpendicular to chip surface connects by contact hole.When making the ground floor conductive channel, not around magnetic core 26, to make beeline channel, but passage 34 is made the almost completely conductive channel of wounded core 26 as shown in Figure 7.This layer conductive channel can be made the phosphorus diffusion method realization of being adopted in the row conductive channel 18 with resembling.Subsequently, on this layer conductive channel, cover a layer insulating 20 again.As shown in Figure 8, second layer conductive channel 36 is deposited on the insulation course 20, and it can be made by employed sputter in the above line conductive channel 30 and lithographic method.Before sputter, insulation course 20 at first is etched out a vertical connecting hole 35 so that little coil channel 34 and 36 connections.Tie point 35 will just in time arrange to allow the afterbody of the little coil conductive channel 34 of ground floor corresponding with the starting point of the little coil conductive channel 36 of the second layer.The same layer insulating 20 that covers repeats to make the 3rd layer of conductive channel 38 with said method, as shown in Figure 9 on the little coil channel 36 of the second layer.Passage 38,34 is drawn array and is connected with pressure welding point 12 (not drawing) with conductor 14.Crucial part is that each conductive channel layer constitutes a single turn on little coil, and each little coil constitutes by " OK " layers 38 of initial 34 an and termination of " row " layer, and wherein belt conductive channel 36 can be according to need and the making multilayer.Notice that all there is slight misalignment in the gap 40 between every adjacent conductive layer, this dislocation arrangement is necessary, it in order to guarantee connecting hole 35 always with the tail end of next conductive layer, the head end of a last conductive layer is corresponding.Equally, little coil also can be with initial method (dopings) realization of making row passage 18.This depends on the requirement of designed device.One of method of mixing in silicon is deposition one deck polysilicon on insulation course 20, makes conductive channel with the mode of lithography guided doping then.After all the little coil layer making except " OK " layer 38 finished, the hole, method generation side 22 with etching made magnetic core 26 with electric plating method again, makes " OK " conductive layer 38 and insulating cover at last to finish structure fabrication.
The benefit of little coil is that each magnetic core can produce the magnetic force more much better than than single-turn circular coil (being directly proportional with the number of turn of little coil), and when particularly being magnetized by the ranks gating at a unit, the magnetization degree of other magnetic cores can be very weak or not be had.
Figure 10, Figure 11 A, Figure 11 B and Figure 11 C have provided out the principle with the little electromagnetic unit of switching gate, and in Figure 10, each unit 41 is connected common power supply 43 together between 45 by tandem tap 37 and 39.Switch 37 can be controlled by the electric signal (curtage) that applies on the lead 30 of being expert at.Switch 39 can be controlled by apply electric signal on column wire 18.And if only if when two tandem taps open, and has electric current to pass through that (being electric current flows out through electromagnetic units 41 to ground 45 from power supply 43 on the unit 41.Switch can be realized with the matrix drive LCD shown in dual-polar triode (bi-polar transistor), the metal oxide semiconductor field effect tube (MOSFET:Metal-Oxide-Semiconductor Field-Effect-Transistor) shown in Figure 11 B or Figure 11 C shown in Figure 11 A.Therefore make switch open or close by the electromotive force that is added in bipolar transistor base stage or metal oxide semiconductor field effect tube grid.Square line of individual pen or the square helical of the multi-turn shown in Figure 11 A shown in Figure 10 and 11B can be made in unit 41.These triodes can be with making with above-mentioned making electromagnetic array similar methods, and can be integrated on the same substrate with conductive channel.In final formation, power supply 43 and ground 45 may be distributed on two conductive layers that separate, and are received respectively on two output terminals of direct supply.Flowing through magnetic core size of current on every side is that supply voltage is divided by the all-in resistance in the circuit (comprising the switch resistance of connection and the resistance of galvanic circle).Backing material is a silicon in the above example, but other materials can be used as substrate as glass, silicide, pottery even plastics.Substrate also can be the material of fine and close or non-densification.Equally, the material of making insulation course 20,28 and 32 also is not limited to above-mentioned exemplifying, and here, they can also be plastics, glass, photoresist, rubber, pottery etc.Conductive layer can be aluminium, gold, tin, copper, titanium, platinum, carbon, semiconductor or their compound.Other structures of little similarly coil and conductive channel also are feasible.Above-mentioned also only is an example by electroplating the method for making magnetic core.Magnetic core can be made by the deposition process in electron beam evaporation, ion sputtering or other microfabrication.Specifically, magnetic core can pass through electron beam evaporation, and ion sputtering or additive method are made a series of Ferrite Material or electromagnetic material by the mode of deposition.But present invention resides in the various single-point gatings of making on the substrate electromagnetic unit that declines.Use this kind chip, biological molecular chemistry reagent and drug molecule all can be transferred control under the action of a magnetic field.
After little electromagnetic array chip manufacturing was finished, thin film be modified or be covered in the surface of insulation course 32 can with chemical method.This tunic is referred to herein as functional layer 42, and it is used to fixedly aglucon molecule.As shown in figure 13, functional layer 42 can be hydrophilic or hydrophobic unimolecular layer, hydrophilic or hydrophobic film, and hydrophilic or hydrophobic gel layer, the compound of poly molecular layer or these materials is as described with Fig. 3 relevant portion.Functional layer 42 also can be made of fine and close or non-dense matter.Can introduce the purpose that has reached immobilization aglucon molecule since specific molecule such as the antibody in functional layer 42, this finally depends on specific aglucon that chip surface will be fixed and analysis or the reaction that will do.These embed in the functional layer and are called as functional group for the molecule that solidifies the aglucon molecule.Being fixed nucleic acid employed aglucon curing materials nitrocellulose or nylon, can be to shift and the functional layer of Northern marking making such as used material such as poly-D-lysine, agarose gel, hydrogel, poly-propionamide gel in shifting at the Southern marking.For ankyrin and polypeptide, other protein moleculars of antibody can be embedded in the functional layer 42 to be used as functional group.
After forming functional layer, the aglucon molecule 44 of being modified or carry (face explanation as follows) by magnetic can be fixed on the functional layer 42 by combining with the difference in functionality group that provides.Figure 13 has described the combination of a kind of direct association reaction such as antibody and antigen, and still, immobilized reactant also not only is confined to this reaction.The immobilization position of aglucon molecule on functional layer 42 can be accurately controlled in the magnetic field that utilizes electromagnetic unit to produce, if promptly in most of the cases single electromagnetic unit 26 is magnetized, the aglucon molecule will be fixed on the functional layer of this top, unit immediately so.As everyone knows, the polarity of electromagnetic field by around this electromagnetic unit around the direction of current decision.Can be polarized into different polarity according to different direction of current (clockwise or counterclockwise) electromagnetic unit.Therefore, when two identical or opposite polarity chrons of the sensed one-tenth of adjacent electromagnetic unit, decision is acted on the magnetic force size on the aglucon molecule of magnetic modified and determines its position that is immobilized by two formed total magnetic fields of the magnetic field superposition that electromagnetic unit produced.And, can adjust the Distribution of Magnetic Field on the chip and change the magnetic force size that acts on the magnetic decorating molecule if by each electromagnetic unit of certain time sequence gating.Make up a fluid pool 46 to be used to hold affinity aglucon molecule, reagent and reactant on chip 10 surfaces, also can carry out the transportation of liquid simultaneously.Figure 12 is the example that the biochip of fluid pool is housed.Fluid pool 46 can be that plastics or other materials are made.The mobile passage that provides of liquid is provided for injection port and outlet 48.(also can use the material of other light-permeables such as glass with silica gel or other materials at the quartzy cover plate 50 of the top seal lastblock of fluid pool 46; Quartz is the material that well is used for ultraviolet detection).Cover plate is for the ease of utilizing light detection method to come the aglucon and the reaction product of pick-up unit inside.If adopt non-optical detection method, fluid pool top 50 just there is no need to use light transmissive material.Example 2: electromagnetic chip with the little electromagnetic unit of planar structure
It below is an example of the electromagnetic chip of the little electromagnetic unit with horizontal structure of the present invention.
What provide in the example 1 is the electromagnetic chip with little electromagnetic unit (vertical cell) of vertical stratification, have horizontal structure little electromagnetic unit (horizontal cell) electromagnetic chip or the electromagnetic chip that has little electromagnetic unit of level and vertical stratification simultaneously all belongs to category of the present invention.The making of these horizontal cells is similar with vertical cell.But consider the size and dimension that horizontal cell is special, make horizontal cell have the character different, be fit to method of the present invention more with vertical cell.
The method of using the present invention to provide can be produced the horizontal cell of various ways.The difference of the horizontal cell of this various ways is the different of the different lead number of turns that center on magnetic core of the size (for example thickness of magnetic core and length) of magnetic core outward.In order to reach residual magnetic field that reduces after the demagnetization and the purpose that increases magnetic pole density, the length that magnetic core should be made is a little, improves the ratio of the length/sectional area of magnetic core.And along with the increase of sectional area, magnetic field intensity also increases.In the process of using electromagnetic chip, before the electric current that reaches capacity, improve the electric signal that applies and also can improve magnetic pole density.(the unit: micron): 200 * 50 * 5200 * 25 * 5200 * 50 * 2200 * 25 * 21600 * 50 * 5400 * 50 * 5400 * 25 * 5400 * 50 * 2400 * 25 * 21600 * 25 * 5 of all size when below having provided design and production technique unit magnetic core
Figure 14 has provided the design drawing of the horizontal cell of being made up of the magnetic core that has centered on 12 circle leads.Magnetic core is of a size of 200 microns * 50 microns, and thickness is about 5 microns.The size of the electric current that the unit applies is 20 milliamperes.Figure 15 is the logarithm equal-value map in the magnetic field of magnetic core in vertical direction 200 microns * 200 micron planars, and the coordinate of figure begins to count from the center of magnetic core.Figure 16 is and the Hm B of Figure 15 in a plane m(Gauss 2/ centimetre) logarithmic graph of (being directly proportional with magnetic field force).Figure 14 B, Figure 14 C and Figure 14 D are the photos of such unit.
Electromagnetic chip with horizontal cell can use 3 inches silicon chips to be raw material, can produce the chip of 1 centimetre of size of 16 1 cm x.16 little electromagnetism horizontally unit of gating separately can be arranged on the chip of 1 centimetre of size of every 1 cm x.Chip with 1 centimetre of size of 1 cm x of 16 horizontal cells is seen Figure 17.Figure 18 then is the silicon chip figure that is manufactured with 16 chip blocks on it.
Figure 19 A be among Figure 14 A along the sectional view of A-A direction, Figure 19 B is along the sectional view of B-B direction among Figure 14 A.Usually, substrate is silicon or glass, and magnetic core is made of CoTaZr, its outer wrapping one deck megohmite insulant, above round gold system lead.The PVT gold thread is to be produced on the silicon chip of oxidized silicon activation by the mode that deposits.Be insulated layer between lead and the magnetic core and separate, insulation course can be photoresist, silicon, monox, silicon nitride and alundum (Al.
Horizontal cell can the vertical unit of use universal method make.Employed megohmite insulant can be divided into 2 types during the production technique unit, so provided 2 kinds of method for makings here.
The photoresist insulation
Step Material Method
Substrate is cleaned and baking Thermal oxidation silicon Scrub
Centering mark deposition Chromium or titanium Sputter
The centering marker mask Positive glue Whirl coating/centering/development
Centering mark etching Ion etching
The photoresist lift off of centering mark is checked Peel off Visual examination
Passivation 0.5 micron SiO 2Or Al 2O 3 Sputter, vapour deposition
Ground floor coil mask Negative glue Whirl coating/centering/development
Ground floor coil deposition 1.0 micron chromium/gold Electron beam evaporation
Ground floor coil etching and inspection Peel off Liftoff/ visual examination/etching
Coil planeization Photoresist Whirl coating/centering/development/baking
The magnetic pole deposition 2.0/5.0 micron CoTzZr Sputter/deposition
Magnetic pole annealing Magnetic vacuum baking/measurement
The magnetic pole mask Positive glue Whirl coating/centering/development
The magnetic pole etching Acid Chemical etching
The magnetic pole photoresist lift off is checked Peel off Visual examination
Magnetic pole passivation mask Photoresist Whirl coating/centering/development/baking
Second layer coil mask Negative glue Whirl coating/centering/development
Second layer coil deposition 1.0 micron chromium/gold Electron beam evaporation
Second layer coil etching and inspection Peel off Liftoff/ visual examination/etching
The coil resistance test Probe card Probe station
Coil passivation mask Photoresist Whirl coating/centering/development/baking
Gold electroplated substrates deposition Titanium/gold Sputter
Gold solder dish mask Positive glue Whirl coating/centering/development
The gold solder dish is electroplated Gold Electroplate
The gold substrate etching Ion etching
The external coating deposition 5～10 microns SiO 2Or Al 2O 3 Sputter
External coating polishes Diamond slurry Polish
Etching external coating exposed pad Ion etching
Final test Probe card Probe station
Insulated by oxide
Step Material Method
Substrate is cleaned and baking Thermal oxidation silicon Scrub
Centering mark deposition Chromium or titanium Sputter
The centering marker mask Positive glue Whirl coating/centering/development
Centering mark etching Ion etching
The photoresist lift off of centering mark is checked Peel off Visual examination
Passivation 0.5 micron SiO 2Or Al 2O 3 Sputter, vapour deposition
Ground floor coil mask Negative glue Whirl coating/centering/development
Ground floor coil deposition 1.0 micron chromium/gold Electron beam evaporation
Ground floor coil etching and inspection Peel off Liftoff/ visual examination/etching
The coil passivation 1.0 micron SiO 2Or Al 2O 3 Sputter
The magnetic pole deposition 2.0/5.0 micron CoTzZr Sputter/deposition
Magnetic pole annealing Magnetic vacuum baking/measurement
The magnetic pole mask Positive glue Whirl coating/centering/development
The magnetic pole etching Acid Chemical etching
The magnetic pole photoresist lift off is checked Peel off Visual examination
Magnetic pole passivation mask 1.0 micron SiO 2Or Al 2O 3 Sputter
The coil channel mask Positive glue Whirl coating/centering/development
The coil channel etching Acid Chemical etching
The coil channel photoresist lift off is checked Peel off Visual examination
Second layer coil mask Negative glue Whirl coating/centering/development
Second layer coil deposition 1.0 micron chromium/gold Electron beam evaporation
Second layer coil etching and inspection Peel off Liftoff/ visual examination/etching
The coil resistance test Probe card Probe station
The coil passivation 1.0 micron SiO 2Or Al 2O 3 Sputter
The pad channel mask Positive glue Whirl coating/centering/development
Pad passage etching Acid Chemical etching
The photoresist lift off of pad passage is checked Peel off Visual examination
Gold electroplated substrates deposition Titanium/gold Sputter
Gold solder dish mask Positive glue Whirl coating/centering/development
The gold solder dish is electroplated Gold Electroplate
The gold substrate etching Ion etching
The external coating deposition 5～10 microns SiO 2 orAl 2O 3 Sputter
External coating polishes Diamond slurry Polish
Etching external coating exposed pad Ion etching
Final test Probe card Probe station
All used some to be used for the mask of photochemistry etching in two kinds of above-mentioned method for makings.During concrete the use, let us choose up to see between magnetic core and the lead and select photoresist and also be to use oxide, mainly will consider the photoluminescent property of resist material itself by some factor decision.Different types of photoresist has different fluorescent characteristics.These materials can and screen to select the photoresist that can be used for fluoroscopic examination by test.Normally used is alundum (Al and monox.
We have made three and have had the chip of horizontal cell and test.Magnetic core is of a size of (unit: micron) in the horizontal cell:
200 * 50 * 5 400 * 50 * 5 1600 * 50 * 5 magnetic field intensitys of measuring in the magnetic core end approximately are about 50% of calculated values, and this may be because the skew that is caused in the manufacturing process.
Device use width is that 5 microns MR head detects, and detects and can carry out on the X-Y-Z platform of manual adjustments at one, and the bearing accuracy of platform is 25 microns.Apply 10 milliamperes electric current on the horizontal cell.Offset point can be regulated by the permanent magnet thin slice of setting in the MR head.The data of measuring can be used the YEW 3033 type instrument records that have the differentiating amplifier module.Wherein the X-axis record is applied to the electric current on the horizontal cell, the signal value on the Y-axis record MR head.
Using before the MR head measures, calibrate with the Helmhotz coil earlier, be aligned in+carry out in/-100 Gausses' the scope.Figure 20 has provided calibration curve.The MR head is installed on the platform in advance, big end to the magnetic core that is positioned at device to be detected.The height setting of detection head is about 50 microns.Applying big electric current on the horizontal cell so that detection head is positioned at peak signal place on the X-Y plane then, i.e. the terminal position of magnetic core, and then carry out subsequently measurement.
The MR head is very sensitive for temperature, so the thermal effect in the measuring process must be taken into account and reduction as far as possible.Apply low frequency (0.25Hz) square wave and help to reduce thermal effect on device, this is that device and detection head system just can reach thermal equilibrium after the several cycles because the amplitude of the electric current that applies is constant.
The relation of the height on measuring magnetic field and electric current, magnetic field and Z direction, the demagnetization situation of device is also tested.The mode of the oscillating current that electric current can be decayed gradually by amplitude decays to zero so that the unit demagnetization, but still has certain residual.
Figure 21 has provided the data of output, is the field intensity-distance map that is of a size of 400 * 50 * 5 magnetic core.Provided the numerical value of component of the Z direction field intensity of the total intensity value calculated and calculating among the figure simultaneously.As seen in the field intensity of magnetic core end mainly along Z-direction.Actual measured value has only about calculated value general, this may be since the final devices of producing on concrete physical dimension and design load exist certain skew to cause.Figure 22 shows that the center of device is approximately 50 MAHs at electric current and reaches capacity (seeing the flex point among the figure), and still along with the increase of electric current, field intensity continues to strengthen, and the magnetic pole density of magnetic core end also continues to increase.Figure 23 has provided the magnetic after effect of device.If the electric signal that directly will apply on the device changes into zero from certain value, will produce the remanent magnetism phenomenon.Left figure applies 20 milliamperes electric current among Figure 23 on the device, removes the remanent magnetism curve of this process of electric current then suddenly.Right figure among Figure 23 applies 20 milliamperes of electric currents on the device, amplitude with electric current decays according to sinusoidal manner then, until the remanent magnetism curve that drops to zero this process, compare with left figure, can learn that the amplitude of using damped system to reduce the electric signal that applies can be so that the remanent magnetism of device be reduced to zero.
Figure 24 A is two kinds of different floor layouts with Figure 24 B, and Figure 24 A is a chip with a plurality of small-sized horizontal cells, and Figure 24 B is a chip with a plurality of big horizontal cells.Figure 25 shows that a plurality of so different chips can be produced on the same silicon chip.
Figure 26 is a structural drawing that is produced on the horizontal cell with square core end structure in the chip surface groove of the present invention.Groove on the chip can be made any suitable size, but the degree of depth preferably and need the particulate or the diameter of aggregation that use approaching among the present invention; The shape of groove also can be various, but preferably circular or oval-shaped.Preferably between 0.5 micron to 10 microns, the width of groove should be between 5 microns to 500 microns, preferably between 20 microns to 200 microns for the degree of depth of groove.Groove can use any suitable method to make, and for example chemistry or laser etching method also can be to use plant equipment to produce groove.In specific operation process, groove can especially be positioned on the magnetic pole on the groove so that particulate attracted on little electromagnetic unit.In cleaning process, the structure of groove can prevent that again particulate is flushed away.Because groove has reduced the distance of electromagnetic unit to chip surface, so groove can also strengthen the intensity of regional internal magnetic field.
The magnetic field intensity of magnetic core end is relevant with the shape of magnetic core end structure.The effect of magnetic core end structure is that the magnetic field intensity with magnetic pole exhales.Figure 27 has provided the structural drawing of multiple magnetic core end.
Below provided a kind of method of production technique electromagnetic unit.At first, need a surface to have the chip of a plurality of leads that are arranged in parallel (for example gold), this can process up by the method for mask and sputter.This chip block will be as the substrate of magnetic core in the horizontal electromagnetic unit of the present invention.On the parallel lead of this layer, deposit one deck megohmite insulant, for example monox earlier.Again at the position deposition magnetic core material that is positioned on this layer megohmite insulant directly over the ground floor parallel wire, the direction of magnetic core should be vertical with the direction of parallel wire below it to small part.The material of magnetic core is CoTzZr preferably, can make arbitrary shape, but preferably makes bar-shapedly, and end is not the shape of directly blocking, but has an inclined-plane, thereby forms outstanding tip.The structure of this end helps to spread the magnetic field of magnetic core, and has relatively long operating distance.On magnetic core, also to cover one deck megohmite insulant again with the zone that originally is not insulated the material covering.
These constitute around the end of the parallel wire of the coil lower floor of magnetic core and can come out by the processing technology of mask etching, for example by the method for acid corrosion the megohmite insulant on the parallel wire end are carved sky, form the vertical cavity that penetrates megohmite insulant.But this cavity can not contact with magnetic core, must have this cavity of megohmite insulant target and magnetic core to be spaced from each other.In cavity, inject the conductor material then, for example golden, so that connect the parallel wire of bottom and the parallel wire on upper strata, to form continuous loop construction.At last, also to cover one deck megohmite insulant again, can also carry out certain processing, for example polishing the surface to obtain smooth surface.In addition, can also use suitable technology at the end of magnetic core, for example the method for mask chemical etching is produced the structure of groove.
Coil can be wired to the lead junction, the lead junction that for example is made of gold.Connecting coil to the lead of lead junction can use suitable material and method (for example mask sputter) to make.The lead junction is in order to connect power supply and coil.
When in coil, applying electric current, just can magnetize magnetic core, thereby produce magnetic field.Magnetic field can use several different methods to measure, and for example uses the MR head.Can select suitable material and size design and making electromagnetic unit according to the purpose of required use electromagnetic unit.Example 3: the method that detects association reaction
Following Example is about electromagnetic chip of the present invention, mainly is to use the electromagnetic chip that has little electromagnetic unit to carry out the detection method of association reaction.
Figure 28 provides according to the present invention to Figure 37, uses electromagnetic biological chip shown in Figure 4 can handle the molecule of chemistry, biology, medicine and other types.These methods may further comprise the steps:
A) construct little electromagnetic array chip 10 of gating separately, as shown in Figure 4.
B) form functional layer 42 at chip surface.This one functional layer is used for the immobilization of aglucon molecule.
As mentioned above, this one functional layer 42 can form by direct chemical modification is carried out on the surface of insulation course 32, or coated one layer of polymeric and forming, and also can form by introducing affinity molecule or active reactive group.This one functional layer can be the compound of hydrophilic or hydrophobic unimolecular layer, hydrophilic or hydrophobic film, functional hydrophilic or hydrophobic glue, polymeric layer, densification or non-tight zone or these materials.
C) the aglucon molecule being carried out magnetic modifies or carries and be fixed on the functional layer 42.
D) on required little electromagnetic unit, produce magnetic field by the electric current of controlling in single lead 18 and 30, the aglucon molecule of modifying or carrying through magnetic will be pulled and be fixed in position specified on the functional layer 42, carry out the different required affinity calmodulin binding domain CaMs of analyzing thereby form on chip.
Can be used for directionally handling and fixing aglucon molecule by diverse ways by applying magnetic field.For example, aglucon molecule 44 can cut linking arm 54 by one and links to each other with paramagnetism magnetic particle 56.Like this, the orientation transportation just can be carried out to the aglucon molecule in the magnetic field that produces by the electromagnetic biological chip, orientation is handled and fixed point discharges.The size of paramagnetism micromagnetism particulate 56 from less than 100 nanometers to all can greater than 100 microns.They can utilize existing technology manufacturing, also can be from buying such as Dynal and the such company of Seradyn.Can cut linking arm 54 and can be light cut, earnestly, enzyme is cut or the cutting of special chemical reaction.It can be covalently bound can cutting linking arm 54 and paramagnetism magnetic particle 56, and perhaps the affinity between the acceptor functional group of functional end-group 52 by can cutting linking arm 54 and magnetic particle 56 is in conjunction with connecting.Be to assemble completely for example below:
Aglucon (44)-can cut linking arm (54)-biotin (52)-streptococcal protein (58)-paramagnetism micromagnetism particulate (56) here, can cut connection between linking arm and paramagnetism micromagnetism particulate lean on be biotin with streptococcal protein between combine and realize.Such molecule assembling can be as any aglucon molecule being carried out the conventional method that paramagnetism micromagnetism particulate is modified, step below the method is used: at first, allow the surface combination (in general, paramagnetism micromagnetism microparticle surfaces has one deck to have carboxyl or amino group) of streptococcal protein and paramagnetism micromagnetism particulate with the method for present maturation.Alternative scheme is directly to buy the paramagnetism micromagnetism particulate that surface coverage has streptococcal protein from producer.Then, preparation " can cut linking arm-biotin " molecular complex.These two steps can be used for the magnetic of any type aglucon molecule and modify.Then, specific aglucon molecule combines with cutting linking arm, as the method by covalent bond.At last, with being coated with the paramagnetism magnetic particle and " aglucon-can cut linking arm-biotin " molecular complex Mixed culture of streptococcal protein, impel the generation of biotin-streptococcal protein association reaction.Whole so intermolecular assembly has just been finished.
In the immobilization of aglucon molecule, the magnetic field that the magnet unit of energising produces applies magnetic field force on paramagnetism micromagnetism particulate 56, and it will make whole minute sub-assemblies contact with the biochip surface of energising magnet unit top.Then, can cut linking arm and be cut, just can be eliminated at magnet unit outage back micromagnetism particulate 56 like this.Sweep-out method can be to utilize fluid flushing or the outside magnetic field force that applies to finish, and stays the aglucon molecule that is fixed on the functional layer 42 at last.
The method of another kind of magnetic carrying aglucon is: paramagnetism micromagnetism particulate and the solution that comprises aglucon are mixed, and rapid again cooling comprises the solia particle 60 (general diameter is less than 1 millimeter) of aglucon and paramagnetism micromagnetism particulate with formation.Solia particle with different specimen preparations can be kept in the refrigerator in order to follow-up use.Use is equipped with the three-dimensional high-precision mechanical arm of the magnetic particle dispensing head 62 of particular design, just such solid micromagnetism particulate can be transported directly to the appointed place on the chip.Behind the precalculated position above solid micromagnetism particulate is transported to the chip appointed area, by control specify little electromagnetic unit electric current so that on the chip magnetic field on the appointed area be better than magnetic field on the dispensing head, just can allow particulate discharge and stationary positioned (Figure 28).Like this, just finished the operation (Figure 29) of appointed area that solid micromagnetism particulate 60 is distributed to the functional layer of chip 10.After solid micromagnetism particulate 60 melted, the aglucon molecule will be fixed on the chip area of appointment (Figure 30) thereupon.Top step has following attendant advantages: use magnetic dispensing head 62 can make the intermolecular cross pollution of aglucon reduce to minimum, dispensing head just need not have been cleaned in so each transportation back.The aglucon molecule is after fixedly the finishing of chip surface, and micromagnetism particulate 56 can clean by externally-applied magnetic field power or liquid above chip it is removed (Figure 31) from chip.
On the chip on each little electromagnetic unit the characteristic dimension of affinity land (rectangle is for long and wide to 5mm at 0.1 μ m; Circle is a diameter) between.The size of calmodulin binding domain CaM is decided by the size of each magnetic core 26 and whether the number of a plurality of magnetic core gatings and gating magnetic core is arranged.The definite size of affinity land also can change by the shape of control function layer 42-and for example, functional layer 42 can be controlled with photoetching and form (opposite is that entire chip is all covered).
E) mark of target sample molecule 62 (for example, with fluorescence 64) be connected with micromagnetism particulate 56.
For the electromagnetic chip that declines with the single-point gating of describing among the present invention comes Controlled Target sample molecule 62, need carry out magnetic earlier to these molecules and modify.Having several different methods to carry out magnetic to target sample molecule modifies.For example, target sample molecule 62 can connect upward paramagnetism magnetic particle 56 by cutting linking arm 54, so just can be by applying magnetic field Controlled Target sample molecule and it being transported to the target area.Can cut the covalent bond of the functional group of terminal functional groups 52 that being connected of linking arm 54 and paramagnetism micromagnetism particulate 56 can be by can cutting linking arm and paramagnetism micromagnetism particulate 56 or acceptor or affinity in conjunction with realizing.For example, connection can be following structure (Figure 32):
Target sample molecule-can cut the such assembly of linking arm-biotin-streptococcal protein-paramagnetic magnetic particle can form by the similar step of top formation " aglucon-paramagnetism micromagnetism particulate " assembly.
F) the target sample molecule 62 that will be connected paramagnetism magnetic particle 56 is placed in the fluid pool 46, again
By controlling magnetic field it is contacted with the aglucon molecule 44 that is fixed on biochip surface.
G) be expert at | in the column unit array, come gating selected with Figure 33 and electric current stream mode shown in Figure 34
In electromagnetic unit
This magnetic field that little electromagnetic unit is produced is alternately turned on and off.Having 13 unit in 25 unit to be energized in Figure 33 then has 12 remaining unit to be energized among Figure 34.Like this, the magnetic field that produces on each little electromagnetic unit can attract target sample molecule 62 and make them shift to the aglucon affinity calmodulin binding domain CaM of appointment.Alternately change the strobe case in magnetic field, each electromagnetic unit just can attract near the target sample molecule it to come to reach the purpose of enrichment from solution.Like this, just can carry out combination and reacted (Figure 35) between target sample molecule 62 and the aglucon molecule 44.
When analyzing on the electromagnetic biological chip that the target sample molecule 62 of magnetic modified is incorporated into, it is STOCHASTIC DIFFUSION (Figure 32) that the motion of target sample molecule 62 begins.To shift to the manipulation of little electromagnetic unit be by realizing as Figure 33 and the magnetic field force that magnetic field produces that alternately opens and closes each unit shown in Figure 34 to target sample molecular orientation afterwards.According to specific analysis requirement, also can target sample molecule 62 directionally be shifted to one or more by these unit of gating optionally by little electromagnetic unit of gating.Under the influence of magnetic field that little electromagnetic unit produced of institute's gating, the target sample molecule 62 of magnetic modified can be shifted to biochip surface rapidly, and affinity reaction (or other reactions) takes place (Figure 35) with the aglucon target sample molecule 44 that is fixed on the designating unit zone.
H) final step, target sample molecule 62 (or their reaction product) separate then with micromagnetism particulate 56 and are eliminated.
Target sample molecule 62 is separated with micromagnetism particulate 56 and can cut cut linking arm 54 (Figure 36,37) between target sample molecule 62 and the micromagnetism particulate 56 by using up methods such as cutting, enzyme cutting, chemical cleavage.The removing of micromagnetism particulate 56 is by applying magnetic field force (this method is not suitable for the situation of having used closed type fluid pond 46) in chip exterior or realizing by the liquid wash that flows through pond 46.
In the top method, aglucon and target sample molecule can be the molecules (learn, materia medica or any other chemical molecular of biological example) of any type.Method described in the present invention can be used for by hybridizing dna molecular, antibody and antigen reactive binding analysis and the drug screening etc. of determining particular sequence (for example drug molecule or potential medicinal compound and special receptor combines).Illustrate, many selective medicinal compounds can be used as the aglucon molecule and be fixed on the precalculated position of functional layer 42.Biological acceptor can separate from cell or produce and do fluorescence labeling with engineered method.Then acceptor is placed the locality on the functional layer 42 that it is combined with candidate compound.After the cleaning, luminous candidate compound is exactly the compound that is hopeful most with the biological acceptor effect on any zone.So the present invention can be used for biochemical reaction, biochemistry detection and diagnostic check.Can finish the complicated macromolecular special organic chemical reactions of assembling in addition.
When above-described method was used for DNA hybridization, behind step h, the dna molecular of non-specific hybridization can be removed in conjunction with condition (as hybridization buffer, temperature etc.) with strict control.Stay the dna molecular that high affinity is arranged with the aglucon molecule at last, they can go to detect with methods such as fluorescence.
When above-described method is used for antibody-antigen-reactive, be right after step h, after cleaning through strict degree, the antibody of non-specific binding or antigen will be eliminated, and the antigen of particular combination or antibody molecule then can be stayed affine land.
When above-described method is used for bioanalysis, the detection of analysis result and quantitative test can realize that for example optical signalling (as light absorption detection or fluoroscopic examination), chemiluminescence or electrochemiluminescence detection, Electrochemical Detection and radio-labeled detect by several detection methods.The fluorescence that optical detection can be produced by the fluorescence 64 that detection laser induces target sample molecule to carry is realized.The method of another kind of optical detection is to send fluorescence and detect with the fluorescence of mark on the induced with laser probe or with fluorescent marker on the secondary antibody that target sample molecular specificity is connected.The fluorescence resonant energy shifts the degree of closeness that also can be used to detect aglucon 44 and target sample molecule 62.Article " Florescence energy transfer dye-labeled primers for DNAsequencing and analysis " (the Proceedings of the National Academy of Sciences that the detailed description that shifts about the fluorescence resonant energy can be delivered people such as Ju, USA, 92:4347-4351,1995) find in.Be with the directed concrete instance of handling of method control dna molecular of the present invention below.
At first, but method having thus described the invention is made little electromagnetic array chip of a single-point gating formula.Surface coverage one deck of chip is used for fixing the high molecular polymer of dna probe.
The adding of paramagnetism micromagnetism particulate is contained in the solution of dna probe, rapidly the potpourri cooling is formed freezing solia particle then.Particulate can be transported to (little electromagnetic unit) on the biochip surface appointed area by the high precision machines people who is equipped with the magnetic dispensing head.Different probe stationary is in different zones.Certainly, each chip different number of probes that can have can be identical with the quantity of little electromagnetic unit on the chip.Little electromagnetic unit on the gating biochip makes the magnetic field of its generation be better than the magnetic field that the magnetic dispensing head is produced, and just can allow the particulate that contains probe be discharged on the functional layer of appointed area on the biochip.After solia particle melted, the dna probe in the liquid just was fixed on the designating unit (zone) of biochip.Then, can remove the micromagnetism particulate by the external magnetic field that is applied to biochip surface, or it be cleaned up by fluid.So just formed the affinity calmodulin binding domain CaM of biochip surface.
Labels targets sample dna molecular (for example, with fluorescein or radiation probe mark) also is connected to the end that light can cut the linking arm molecule.Another end at linking arm is a biotin molecule.The streptococcal protein molecule is fixed on the surface of magnetic micromagnetism particulate.Like this, just can be connected to by the interaction of biotin-streptococcal protein on the magnetic micromagnetism particulate with target sample dna molecular after the paramagnetism micromagnetism particulate that is coated with streptococcal protein mixes when the solution that contains target sample DNA-linking arm-biotin composite.
DNA target sample molecule-light can cut linking arm-biotin-streptococcal protein-magnetic micromagnetism particulate.
Then, the solution that will contain the target sample dna molecular modified of being magnetic is introduced in the fluid pool on the biochip.The alternately gating of little electromagnetic unit makes each units alternately on the chip produce magnetic field.The target sample dna molecular of modifying through paramagnetism micromagnetism particulate just moves to the dna probe molecule that is fixed on the chip surface thereupon.Because all electromagnetic units all can be by gating, so target sample dna molecular just is directed contacting with all dna probes.Target sample dna molecular just can carry out hybridization reaction at affinity calmodulin binding domain CaM and probe molecule under the hybridization conditions of selecting in advance like this.Can both detect by fluorescence, luminous or radiation with any probe of target sample dna molecule hybridize, concrete mode depends on the label that uses on target sample molecule.Can do screening at a plurality of dna probes to given DNA target sample molecule apace in this way, it is fast to reach analysis speed, the requirement that automaticity is high.If micromagnetism particulate interference detection, then they can be removed from target sample dna molecular, for example cut the operation that linking arm comes DNA isolation and magnetic particle adopting light to cut just can to reach this using up by rayed under the situation of linking arm with 250nm-750nm.Magnetic particle can be removed it by the magnetic force or the washing fluid that add from the conversion zone on the chip freely subsequently.In addition, can also allow chip be in " hot melt dissociates " temperature to remove the target sample DNA of having hybridized and to reuse.
The inventor believes that above-mentioned example shows the method for optimizing of using this invention.Yet described parameter such as size, material, geometric configuration, method, experiment rules, temperature, concentration and time are not regarded as limiting of the invention.Except that the part of making claim, those skilled in the art can all belong to the coverage of this patent at tangible substitute present or that understand later on.Therefore the special scheme of describing above this claim is understood to include, and those schemes that can substitute significantly.Described specific embodiment only is in order to reach for example purpose, and should not think that the present invention just only is confined to this.Therefore, should be appreciated that within the scope of the appended claims that except that the situation of the special description of doing in this place, the present invention is suitable in the same old way.
Example 4: the usage level unit carries out the capable ripple of magnetic force
Row ripple magnetophoresis can realize by the electromagnetic unit of various ways, described belowly is to use horizontal cell of the present invention to go the ripple magnetophoresis.
In order to go the ripple magnetophoresis, should have reaction tank on the electromagnetic chip as shown in Figure 24 A and 24B.On the functional layer on the electromagnetic unit, 7 kinds of lymphoma cells of specific recognition and a kind of white blood cell, for example antibody of B cell have been fixed.In addition, prepared these antibody simultaneously and be mixed into solution state, rather than be fixed on chip surface.Sample (comprising blood) and pan coating the magnetic particle of antibody mix the antigen combination that antibody can be special with cell surface.According to concrete needs, after this step operation, can also in solution, add reagent to handle sample.For example, interpolation can splitting erythrocyte solution, better this solution is splitting erythrocyte only, the basic step influences white blood cell, best situation is that influence (can be referring to U.S. Patent application " Compositions and Methods for Separation of Moieties on Chips " or do not influence substantially to dielectrophoresis for this solution, on October 10th, 2000 submitted, Patent Office's numbering: 09/686,737).Sample, sample solution and additional solution, damping fluid, reagent can add reaction tank in any mode easily, for example shift by transfer pipet, injector to inject, utilize gravity to introduce by conduit (for example polyethylene catheter or the like).Generally, sample, sample solution and other solution, damping fluid, reagent add reaction tank in the mode of continuous flow, such continuous flow can be injected into or pump into reaction tank from least one inlet, then, do not contain sample component and other liquid and flow out reaction tank from least one outlet.
Before sample adds reaction tank, can in sample, add sample solution earlier.Before sample-sample solution mixed liquor is added reaction tank, sample and sample solution jointly incubation from less than 1 second to several hrs time arbitrarily among several days.In the conduit that the mixing of sample and sample solution can occur in reaction tank is communicated with.Also can earlier sample be added reaction tank, then sample solution be added reaction tank again; Or earlier sample solution is added reaction tank, then sample is added reaction tank again.
Wherein bond such as the magnetic particle that uses in the present invention can provide together or separately with sample solution.If bond provides separately, so bond can be before sample solution adds sample, add simultaneously or afterwards.
Sample solution can add sample or comprise in the reaction tank of electromagnetic chip and mix before sample deposition is to the electromagnetic chip.Sample-sample solution mixed liquor is being fed before reaction tank separates, sample and sample solution can leave standstill a period of time jointly, from can less than 1 second to several hours even several days.The mixed process of sample and sample solution can occur in the conduit that feeds reaction tank.Perhaps, can earlier sample be joined in the reaction tank, again sample solution be added reaction tank; Also can earlier sample solution be joined in the reaction tank, again sample be added reaction tank.
Usually, magnetic particle should be added sample and carry out leaving standstill a period of time before the magnetic particle separation, during this period of time can be from extremely several hours even several days a few minutes.Can the selectivity born of the same parents separate erythrocytic sample solution add sample before, simultaneously or in sample, add magnetic particle afterwards.
Sample, sample solution and additional solution, damping fluid, reagent can add reaction tank in any mode easily, for example shift by transfer pipet, injector to inject, utilize gravity to introduce by conduit (for example polyethylene catheter or the like).Generally, sample, sample solution and other solution, damping fluid, reagent add reaction tank in the mode of continuous flow, such continuous flow can be injected into or pump into reaction tank from least one inlet, then, do not contain sample component and other liquid and flow out reaction tank from least one outlet.
Can be by one or a plurality of conduit feeds one or more additive reagents in sample, as particulate, but this is not of the present invention necessarily requiring.For example, can add sample as particulate, leave standstill jointly after a period of time, again sample be joined in the reaction tank earlier with one or more additive reagents.Perhaps, particulate can contact with sample by the conduit that links to each other with reaction tank, and like this, particulate can mix with sample in sample flows into the process of reaction tank mutually.Also can be when sample be fed reaction tank or afterwards, with additive reagent such as particulate by one or a plurality of conduit feeds reaction tank.Use if desired more than a kind of additive reagent, then they can or add sample respectively according to the above-mentioned method while.
Sample can be gone up the random ripple magnetophoresis of going at chip (Figure 38 A, 38B and 38C) by using particulate switch (Figure 39 A).Have leukocytic particulate and flow through and be fixed with the zone of special antibody, if the identification white blood cell that fixing antibody can be special on the functional layer, white blood cell-particle complex will be combined in zone specific on the chip.First-selected fixing antibody is those antibody that can discern lymphocyte (for example T cell, B cell, macrophage, neutrophil(e) cell etc.).The distribution situation of the cell of these kinds can reflect the health status of the biology of sampling in the sample, for example, can judge whether biology exists lymthoma, judgement to biosome own type immunological diseases (comprise that HIV infects, acquired immunodeficiency disease with and relevant disease) the treatment situation, investigate the situation of other disease of biosome, comprise septicemia.These treatment of diseases situations can make in this way monitors at any time, for example monitors the variation of CD4+ cell quantity in HIV cases of infection therapeutic process.
The antibody of other kind also can use, and for example at the antibody of cancer antigen, particularly those are positioned at the antigen on cancer cell (as breast cancer cell, lung carcinoma cell, prostate gland cancer cell or the like) surface.By method of the present invention, detect these cells and whether be present in the chip surface certain location, can diagnose whether suffer from these diseases, or detect the advancing of disease situation.The antibody of other directed toward bacteria, parasite, virus or prion also can use.
Reaction tank can clean earlier, the unconjugated material of flush away, and then add and to have fluorescently-labeled antibody and detect.Used 8 kinds of antibody in this example, every kind can be used identical or different antibody to detect.The antibody of the tape label that adds is aimed at the antibody that is fixed on the functional layer.The antibody of fixing antibody, cell and tape label forms the compound of sandwich structure.
Reaction tank can clean earlier, and the unconjugated material of flush away adopts the electromagnetic wave of certain wavelengths to excite according to employed mark then.Signal on the chip can use CCD equipment to receive.Detect image and can use suitably, for example use disk or CD by storing.Also can use the MR head to determine the position of magnetic particle on chip.
Fluoroscopic examination figure on the chip can show sample in leukocytic information, particularly whether have lymphoma cell.Leukocytic distribution per sample can be to the lymphoma cell in the sample classify (seeing Figure 40).
All titles all are the convenience for the reader in the literary composition, do not play content under the restriction title.

Claims (56)

  1. But 1, a kind of have a decline electromagnetic chip of electromagnetic unit of single-point gating, comprising:
    A) substrate;
    B) be positioned at on-chip a plurality of little electromagnetic unit, can make each unit produce magnetic field by applying electric current;
    C), make it produce the method in magnetic field in order to any one or a plurality of unit in a plurality of little electromagnetic units of gating;
    Having an above-mentioned little electromagnetic unit at least is planar structure.
  2. 2, according to the electromagnetic chip of claim 1, wherein each little electromagnetic unit comprises:
    A) be arranged on an on-chip magnetic core;
    B) be used for the method for wounded core conduction current;
    Wherein above-mentioned magnetic core is meant magnetic core or the core that can be magnetized.
  3. 3, according to the electromagnetic chip of claim 2, magnetic core wherein is that ferrimagnet or ferrous section bar material are made.
  4. 4, according to the electromagnetic chip of claim 2, the device that wherein is used for the wounded core conduction current is the single-turn or multi-turn electrical lead of wounded core.
  5. 5, according to the electromagnetic chip of claim 4, wherein the electrical lead shape of wounded core can be circular, square, oval, triangle, spirality and right angle spirality, and the loop that these leads form can all be in a plane, also can be in different planes.
  6. 6, according to the electromagnetic chip of claim 4, be variable wherein from the size and Orientation that is applied to the electric current on little electromagnetic unit that current source transmits.
  7. 7, according to the electromagnetic chip of claim 1, wherein the device of any little electromagnetic unit of gating comprises:
    Be connected the switch between current source and each the little electromagnetic unit, this switch can be controlled open/close state and thus the strobe case of little electromagnetic unit be controlled.
  8. 8, according to the electromagnetic chip of claim 7, wherein said switch can be the switch of machinery, also can be the switch of electronics, also can be the assembly of above-mentioned two kinds of form switches.
  9. 9, according to the electromagnetic chip of claim 1, wherein little electromagnetic unit is arranged in substrate surface by the orderly repeat patterns of rule, and the adjacent cells spacing equates.
  10. 10, according to the electromagnetic chip of claim 1, wherein the size of little electromagnetic unit is between 0.1 micron to 1 centimetre.
  11. 11, according to the electromagnetic chip of claim 1, comprise at least one one functional layer, can be used for the fixing of at least a aglucon or entity molecule.
  12. 12, electromagnetic chip according to claim 11, it is characterized in that, the described functional layer that is used for fixing the aglucon molecule can be hydrophilic unimolecular layer, hydrophilic unimolecular layer with functional group, hydrophobic unimolecular layer has the hydrophobic unimolecular layer of functional group, hydrophilic film, hydrophilic film with functional group, hydrophobic film has the hydrophobic film of functional group, hydrophilic gel layer, hydrophilic gel layer with functional group, the organophilic gel layer has the organophilic gel layer of functional group, non-dense material, non-dense material with functional group, dense material and dense material with functional group.
  13. According to the electromagnetic chip of claim 12, it is characterized in that 13, described functional group can be aldehyde radical, carbodiimide, succinimide ester, antibody, acceptor or lectin.
  14. 14,, comprise a kind of entity molecule or aglucon at least and be fixed on the described functional layer according to the electromagnetic chip of claim 11.
  15. 15, according to the electromagnetic chip of claim 14, it is characterized in that described entity molecule or the aglucon molecule comprises oligonucleotide, DNA, RNA, polypeptide, albumen, carbohydrates, lipid, eukaryotic, prokaryotic, prion, virus, parasite, antibody, agglutinin or acceptor.
  16. 16, a kind of electromagnetic chip consists of:
    A) substrate;
    B) one or a plurality of little electromagnetic unit, the consisting of of little electromagnetic unit:
    I) magnetic core;
    Ii) center on the lead loop of magnetic core;
    Wherein said magnetic core is horizontal direction on electromagnetic chip.
  17. According to the electromagnetic chip of claim 16, it is characterized in that 17, described little electromagnetic unit is arranged in the form of array.
  18. 18, according to the electromagnetic chip of claim 16, it is characterized in that having a gating separately in described little electromagnetic unit at least.
  19. 19, according to the electromagnetic chip of claim 16, it is characterized in that whole gating separately in described little electromagnetic unit.
  20. According to the electromagnetic chip of claim 16, it is characterized in that 20, the described array of being made up of a plurality of little electromagnetic units is parallel to each other.
  21. According to the electromagnetic chip of claim 20, it is characterized in that 21, the described array of being made up of a plurality of little electromagnetic units comprises a particulate switch.
  22. According to the electromagnetic chip of claim 20, it is characterized in that 22, the described array of being made up of a plurality of little electromagnetic units can be used as a capable ripple magnetophoresis device.
  23. According to the electromagnetic chip of claim 16, it is characterized in that 23, what apply on described little electromagnetic unit is sinusoidal signal.
  24. 24, a kind of method of making little electromagnetic unit, step is:
    A) plane;
    B) have a plurality of conductor materials that are arranged in parallel on the plane;
    C) deposition ground floor megohmite insulant on the parallel conductor material of this layer;
    D) again at the position deposition magnetic core material that is positioned on this layer megohmite insulant directly over the ground floor parallel conductor material;
    E) deposition second layer megohmite insulant on magnetic core;
    F) expose the end of formation around the parallel conductor material of the coil lower floor of magnetic core;
    G) add the upper conductor material to form coil;
    H) the three-layer insulated material of deposition on described coil.
  25. 25, a kind of little electromagnetic unit is characterized in that, is to use the described method of claim 24 to make.
  26. 26, the array of being made up of little electromagnetic unit is characterized in that, is to use the described method of claim 24 to make.
  27. 27, an electromagnetic chip contains the array of being made up of little electromagnetic unit, it is characterized in that, is to use the described method of claim 24 to make.
  28. 28, a kind of little electromagnetic unit, contain:
    A) the ground floor lead that is separated by ground floor megohmite insulant and magnetic core;
    B) one deck conductor material is connected with above-mentioned ground floor lead to form the outer coil of magnetic core; Make coil and magnetic core insulate by megohmite insulant; When coil applies electric current, produce magnetic field in the magnetic core.
  29. 29, a kind of array of being made up of little electromagnetic unit is characterized in that, wherein contains a described little electromagnetic unit of claim 28 at least.
  30. 30, a kind of electromagnetic chip is characterized in that, wherein contains a described little electromagnetic unit of claim 28 at least.
  31. 31, a kind of capable ripple magnetophoresis device is characterized in that, wherein contains two or the described little electromagnetic unit of a plurality of claim 28.
  32. 32, a kind of particulate switch is characterized in that, wherein contains two or the described little electromagnetic unit of a plurality of claim 28.
  33. 33, a kind of little electromagnetic unit comprises:
    A) magnetic core;
    B) be insulated the ground floor lead that material and magnetic core separate;
    C) the conductor material connects above-mentioned ground floor lead and second layer lead;
    Described ground floor lead, conductor material and second layer lead are formed the outer coil of magnetic core;
    Magnetic core and coil insulate by megohmite insulant.
  34. 34,, comprise one deck megohmite insulant at top according to little electromagnetic unit of claim 33.
  35. 35, according to little electromagnetic unit of claim 33, magnetic core comprises the stretched out structure that at least one is terminal.
  36. 36,, comprise the structure of at least one groove according to little electromagnetic unit of claim 33.
  37. 37, a kind of array of being made up of little electromagnetic unit is characterized in that, comprises at least one little electromagnetic unit described in 33.
  38. 38, a kind of electromagnetic chip is characterized in that, comprises the little electromagnetic unit described at least one claim 33.
  39. 39, a kind of capable ripple magnetophoresis device is characterized in that, comprises the little electromagnetic unit described in two or a plurality of claim 33.
  40. 40, a kind of particulate switch is characterized in that, comprises the little electromagnetic unit described in two or a plurality of claim 33.
  41. 41, according to the electromagnetic chip of claim 33, wherein the material of ground floor insulation course can be any one in the materials such as monox, silicon nitride, plastics, glass, pottery, photoresist, rubber.
  42. 42,, comprise one deck substrate according to the electromagnetic chip of claim 33.
  43. According to the electromagnetic chip of claim 42, it is characterized in that 43, the material of substrate can be any one in silicon, glass, pottery, monox or the plastics.
  44. According to the electromagnetic chip of claim 33, it is characterized in that 44, the material of lead can be any one in the materials such as aluminium, gold, silver, tin, copper, platinum, palladium, carbon or semiconductor.
  45. 45, according to the electromagnetic chip of claim 33, it is characterized in that, also comprise an one functional layer.
  46. 46, electromagnetic chip according to claim 45, it is characterized in that, the described functional layer that is used for fixing the aglucon molecule can be hydrophilic unimolecular layer, hydrophilic unimolecular layer with functional group, hydrophobic unimolecular layer has the hydrophobic unimolecular layer of functional group, hydrophilic film, hydrophilic film with functional group, hydrophobic film has the hydrophobic film of functional group, hydrophilic gel layer, hydrophilic gel layer with functional group, the organophilic gel layer has the organophilic gel layer of functional group, non-dense material, non-dense material with functional group, dense material and dense material with functional group.
  47. According to the electromagnetic chip of claim 33, it is characterized in that 47, functional group can be acetaldehyde, carbodiimide, succinimide ester, antibody, acceptor or lectin.
  48. 48, according to the electromagnetic chip of claim 33, it is characterized in that, comprise that also one makes liquid and array fluid in contact pond.
  49. 49, a kind of method of operating magnetic particulate comprises:
    A) provide a kind of electromagnetic chip, but this chip contains little electromagnetic unit of a plurality of single-point gatings;
    B) sample that will contain magnetic particle is added to the surface of chip;
    C) change the electric current be added on each little electromagnetic unit changing the Distribution of Magnetic Field on the chip surface, thereby change the magnetic field force that is added on the magnetic particle.
  50. According to the method for claim 49, it is characterized in that 50, comprise at least one entity molecule on the described magnetic particle, this entity molecule is connected on the magnetic particle.
  51. According to the method for claim 49, it is characterized in that 51, described connection realizes by linking arm, covalent bond or biological compatible reaction.
  52. 52, according to the method for claim 49, it is characterized in that described entity molecule is oligonucleotide, DNA, RNA, polypeptide, albumen, carbohydrates, lipid, eukaryotic, prokaryotic, prion, virus, parasite, antibody, agglutinin or acceptor.
  53. 53, according to the method for claim 49, described electromagnetic chip comprises a magnetophoresis device.
  54. 54, according to the method for claim 49, described electromagnetic chip comprises a particulate switch.
  55. 55, according to the method for claim 49, the magnetic core on described little electromagnetic unit contains an end structure at least.
  56. 56,, contain the structure of groove on the described electromagnetic chip according to the method for claim 49.
CN 01136350 2000-10-10 2001-10-10 Planar microelectromagnetic unit array chip capable of being gated separately Expired - Fee Related CN100476431C (en)

Applications Claiming Priority (2)

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US09/685,410 2000-10-10
US09/685,410 US6716642B1 (en) 1999-03-15 2000-10-10 Individually addressable micro-electromagnetic unit array chips in horizontal configurations

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CN1348103A true CN1348103A (en) 2002-05-08
CN100476431C CN100476431C (en) 2009-04-08

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