CN101710087B - Micron-nano-sized single particle impedance spectroscopy measurement chip and measuring method - Google Patents
Micron-nano-sized single particle impedance spectroscopy measurement chip and measuring method Download PDFInfo
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- CN101710087B CN101710087B CN200910213589A CN200910213589A CN101710087B CN 101710087 B CN101710087 B CN 101710087B CN 200910213589 A CN200910213589 A CN 200910213589A CN 200910213589 A CN200910213589 A CN 200910213589A CN 101710087 B CN101710087 B CN 101710087B
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Abstract
The invention provides a micron-nano-sized single particle impedance spectroscopy measurement chip and a measuring method, relating to the surface technology of micron-nano-sized single particles (including cells), in particular to the impedance spectroscopy measurement technology of the micron-nano-sized particles; in the measurement chip, an upper cover board (01) is a transparent material, a transparent electricity-conduction film layer (02) is arranged on the lower surface of an upper cover board (01), an electricity-conduction film (08) is arranged on a chip lower substrate (09), a photoconductive layer (07) is arranged on the electricity-conduction film (08) at the lower layer, an insulating layer (06) is arranged on the photoconductive layer (07), a single particle clamping block (05) is arranged on the insulating layer (06), an interlayer (04) is arranged between the transparent electricity-conduction film layer (02) at the upper layer and the electricity-conduction film (08) at the lower layer, and an injection port (03) is arranged at one side of the upper cover board (01); the single particle impedance spectroscopy measuring method comprises the following steps: a voltage signal for particle control is switched on, a target particle is captured in a particle swarm area (11) by utilizing a micrographic light pattern (12) and is conveyed to the groove between the single particle clamping blocks (051 and 052), and then the particle control voltage is switched off, the impedance spectroscopy measurement voltage is switched on, the impedance spectroscopy measurement of the single particle is carried out by combining a dynamic signal analyzer.
Description
Technical field
The present invention relates to the impedance spectrum measuring technique of micro-nano the single-particle characterization technique of (containing cell), particularly micro-and nano-particles.
Background technology
The impedance spectrum measurement is to apply alternating voltage through the two ends at sample, through signal analyzer, draws the variation relation of the impedance of the particle colony in the sample with frequency then.The impedance spectrum measurement is a kind of important method of the electrical characteristics of research cell or other micro-and nano-particles, can realize the sign to micro-nano particle colony.Electrical impedance test and analysis have all obtained in fields such as drug screening, medical diagnosis on disease and treatment, Food Inspection, environmental monitorings using widely.Impedance spectrum measurement towards biological sample is called bio-impedance technology (bio-impedancetechnology), and it has fast, simple, the easy-operating characteristics of electrode structure, progressively becomes effective analysis tool in the biomedical research.
Present impedance spectrum measuring technique towards be particle colony, to the control of single particle with measure powerlessly, can't realize certain the specific single-particle in the particle colony is measured.And under many circumstances; Research to the impedance spectrum of single particle (containing unicellular) is all necessary and significant: can pick out the unique electrical property of very small particles in the same particle colony; Perhaps can be careful tell the difference between any two particles in the same particle colony, perhaps realize the impedance operator of certain specific single-particle is studied.
Therefore, on the basis of inheriting above-mentioned impedance spectrum basic principle of measurement, how to realize that it is the key issue that solves above-mentioned limitation that certain specific single-particle in the sample colony is carried out impedance measurement.
Summary of the invention
Technical matters: the purpose of this invention is to provide micro-nano single-particle the impedance spectrum measured chip and the measuring method of (containing unicellular), be difficult to realize the problem that impedance spectrum is measured of carrying out to certain specific in sample single-particle to solve present impedance spectrum measuring technique.
Technical scheme: the invention provides the measured chip and the measuring method of a kind of micro-nano single-particle (containing unicellular) impedance spectrum, can choose certain single-particle in the same particle colony arbitrarily and in same chip, accomplish impedance spectrum and measure.The technical scheme that the present invention adopts is:
Sized single particle impedance spectroscopy measured chip provided by the invention comprises upper cover plate, upper strata transparent conductive film, injection port, wall, single-particle grip block, insulation course, photoconductive layer, lower conductive film, following substrate and chip internal cavity.Upper cover plate is a transparent insulation material, can select glass or organic glass to make; Wall is made by insulating material, and between upper strata transparent conductive film and lower conductive film, is used for the inner cavity of supporting chip; Substrate is provided with conductive film under the chip, on lower conductive film, is provided with photoconductive layer, on conductance layer, is provided with insulation course, on insulation course, is provided with the single-particle grip block, and injection port is positioned at a side of upper cover plate; The single-particle grip block comprises two sub-block of pairing, and the sub-piece of a pair of clamping is good conductor.When the impedance spectrum of the parallel a plurality of particles of measurement of needs, can have manyly to the sub-piece of clamping, every pair of sub-piece of clamping is used for the measurement of a particle.Photoconductive layer is by the material with photoconductive property, and following substrate is an insulating material.
Between upper strata transparent conductive film and lower conductive film, apply and be used for the alternating voltage that particle is controlled; Clamping is applied the alternating voltage that is used for the particle impedance measurement between the sub-piece at each.When being used for alternating voltage that particle controls and connecting; Use miniature light pattern direct-writing device to generate small dynamic light pattern (light pattern can real time kinematics); And with miniature light pattern from chip below or the top be projected on the photoconductive layer; Because the area pellucida on the photoconductive layer is different with the conductivity of dark space, make to form inhomogeneous field in the cavity, and then the formation dielectrophoretic force.Above-mentioned miniature light pattern direct-writing device obtains picture signal from computing machine and forms small light pattern and then project the tiny area on the chip through DMD (DMD) drive plate generation dynamic light pattern and through the lens optical path unit.Through to the projection of digital micro-mirror control, the aperture pattern is caught certain single particle in the population in the sample, and, particle is held between the sub-piece of a pair of clamping in the groove of this PARTICLE TRANSPORT FROM in the middle of the grip block.After applying electric signal on these two the sub-pieces of conduction clamping; Electric current can be through tested particle (particle that promptly grasps); Signal analyzer through the outside is to analyzing through the electric signal at tested particle two ends then; Through conversion of signals, obtain the impedance (or admittance) of the tested particle in the certain frequency scope.Catch and the transport process of single particle can be observed and record the motion conditions of real-time monitoring objective particle through the micro image collection device.Image collecting device comprises microscope, ccd video camera and IMAQ converting unit, and its output terminal is connected with computing machine.
Single-particle provided by the invention (containing unicellular) impedance spectrum measuring method is:
Step 1: between upper strata transparent conductive film and lower conductive film, apply and be used for the ac voltage signal that particle is controlled;
Step 2: use miniature light pattern direct-writing device to generate miniature aperture pattern (zone beyond the light pattern is the dark space), and be projected on the photoconductive layer of chip, move to the zone that population belongs to then and with miniature aperture, catch the target single-particle;
Step 3: the direction of the particle of catching to grip block transported, and intended particle is sandwiched in the groove between the sub-piece of a pair of clamping.
Step 4: be used for the voltage signal that particle controls and close with aforementioned; Between the sub-piece of a pair of clamping, apply the ac voltage signal that is used for impedance measurement then; Make electric signal pass through tested single-particle; The frequency in conditioning signal source in predetermined scope, and gather the information such as amplitude, phase place of the sine voltage signal on the sub-piece of a pair of clamping in real time by signal analyzer and then through calculating the resistance value of tested particle.
Beneficial effect: the present invention proposes micro-nano single-particle (containing cell) impedance spectrum measured chip and measuring method; On the basis of the advantage that the inheriting tradition impedance spectrum is measured; Through the sub-piece of the clamping of photoconductive layer and pairing being integrated among the chip, realize the measurement of single particle (comprising cell) impedance spectrum.The present invention catches the single target particle through dynamic miniature light pattern and intended particle is transported between the sub-piece of a pair of clamping; Manipulation process has very high flexibility and reconfigurability, helps realizing the selective measurement of the Different Individual particle in the same particle colony.The sub-block structure of the clamping of pairing provided by the invention has advantage simple in structure, that be convenient to make.Impedance spectrum measuring method step provided by the invention is few, simple operation, quick.
Description of drawings
Fig. 1 is the structural representation of the sized single particle impedance spectroscopy measured chip in the embodiment of the invention;
Fig. 2 is the vertical view that the sized single particle impedance spectroscopy in the embodiment of the invention is measured the examination chip internal.
Have among the above figure:
The sub-piece of the sub-piece of first clamping 051, second clamping 052, particle colony aggregation zone 11, catch the particle position 131 under aperture 12, the trapped state, the particle position 132 under the clamp position.
Embodiment
The embodiment of sized single particle impedance spectroscopy measured chip provided by the invention comprises upper cover plate 01, upper strata transparent conductive film 02, injection port 03, wall 04, single-particle grip block 05, insulation course 06, photoconductive layer 07, lower conductive film 08, substrate 09 down, chip internal cavity 10 referring to Fig. 1 and Fig. 2.Upper cover plate 01 is a transparent material, can select glass or organic glass to make; The material of transparent conductive film layer 02 can be selected indium and tin oxide film for use; Wall 04 is made (for example can select the SU-8 photoresist for use) by insulating material, is used for the inner cavity 10 of supporting chip; Single-particle grip block 05 comprises sub-piece 051 of first clamping and the sub-piece 052 of second clamping; The sub-piece of clamping (051 and 052) is good conductor, can select material such as gold, silver, aluminium for use; Insulation course 06 can select for use silicon nitride material to make; Photoconductive layer 07 can be selected amorphous silicon hydride for use; Lower conductive film 08 can be transparent also can be opaque; Following substrate 09 can be selected glass or silicon chip.
In the present embodiment, between upper strata transparent conductive film 02 and lower conductive film 08, apply first via alternating voltage (being used for particle controls); Between sub-piece 051 of first clamping and the sub-piece 052 of second clamping, apply the second road alternating voltage (being used for impedance spectrum measures).
The embodiment of sized single particle impedance spectroscopy measuring method provided by the invention is referring to Fig. 2.When first via alternating voltage (being used for particle controls) is connected; Use miniature light pattern direct-writing device to generate small dynamic light pattern; And with miniature light pattern from chip below or the top be projected on the photoconductive layer 07; Because the area pellucida on the photoconductive layer 07 is different with the conductivity of dark space, make to form inhomogeneous field in the cavity, and then the formation dielectrophoretic force.Above-mentioned miniature light pattern direct-writing device obtains picture signal from computing machine and forms small light pattern and then project the tiny area on the chip through DMD (DMD) drive plate generation dynamic light pattern and through the lens optical path unit.Through projection control to digital micro-mirror; The position 131 that can use aperture pattern 12 certain single particle place in population 11 is with this particle trapping; And, particle is held between sub-piece 051 of first clamping and the sub-piece 052 of second clamping with the clip position 132 of this PARTICLE TRANSPORT FROM to particle.After applying electric signal on these two the sub-pieces of conduction clamping; Electric current can be through tested particle (particle that promptly grasps); Through the dynamic signal analyzer of outside the electric signal through intended particle is analyzed then; Through its inner conversion of signals, obtain the impedance (or admittance) of the tested particle in the certain frequency scope.Catch and the transport process of single particle (comprising cell) can be observed and record the motion conditions of real-time monitoring objective particle through the micro image collection device.Image collecting device comprises microscope, ccd video camera and IMAQ converting unit, and its output terminal is connected with computing machine.
The concrete steps of the embodiment of the measuring method of sized single particle impedance spectroscopy provided by the invention are following:
Step 1: between upper strata transparent conductive film 02 and lower conductive film 08, apply and be used for the ac voltage signal that particle is controlled;
Step 2: use miniature light pattern direct-writing device to generate miniature aperture pattern (zone beyond the light pattern is the dark space), and be projected on the photoconductive layer 07 of chip, then miniature aperture 12 is moved to the zone 11 at population place, catch the single target particle;
Step 3: the direction of the particle of catching to grip block 05 moved, and intended particle is sandwiched in the V-type groove between sub-piece 051 of first clamping and the sub-piece 052 of second clamping.
Step 4: will be used for particle and control ac voltage signal and close.On first grip block 051 and second grip block 052, apply the ac voltage signal that is used for impedance measurement then; Make the frequency in conditioning signal source in predetermined scope, and gather in real time that first grip block 051 and second adds information such as the amplitude of holding the sine voltage signal on the piece 052, phase place and then through calculating the resistance value of tested particle through signal analyzer.
Claims (2)
1. a micron-nano-sized single particle impedance spectroscopy measured chip is characterized in that this impedance spectrum measured chip comprises upper cover plate (01), upper strata transparent conductive film (02), injection port (03), wall (04), single-particle grip block (05), insulation course (06), photoconductive layer (07), lower conductive film (08), substrate (09) down, chip internal cavity (10); Upper cover plate (01) is a transparent material; Upper strata transparent conductive film layer (02) is positioned at the lower surface of upper cover plate (01); Substrate under the chip (09) is provided with lower conductive film (08); On lower conductive film (08), be provided with photoconductive layer (07), on photoconductive layer (07), be provided with insulation course (06), on insulation course (06), be provided with single-particle grip block (05); Wall (04) is positioned between upper strata transparent conductive film (02) and the lower conductive film (08) and is positioned at insulation course (06), photoconductive layer (07) both sides, and injection port (03) is positioned on the upper cover plate (01);
Single-particle grip block (05) in the described chip comprises the first sub-grip block (051) and the second sub-grip block (052) of two pairings, and has the groove structure between two grip blocks and be used for the target single-particle is grasped;
Described single-particle grip block (05) is a good conductor, and single-particle grip block (05) can have one or more, and described target single-particle comprises individual cells and other micro-nano biomone.
2. measuring method that is used for the described micron-nano-sized single particle impedance spectroscopy measured chip of claim 1 is characterized in that this method may further comprise the steps:
Step 1: between upper strata transparent conductive film (02) and lower conductive film (08), apply and be used for the voltage signal that particle is controlled;
Step 2: use miniature light pattern direct-writing device to generate miniature aperture pattern (12), and be projected on the photoconductive layer (07) of chip, then miniature aperture is moved to the zone (11) at population place, catch the single target particle;
Step 3: the direction of the single-particle of catching to grip block (05) transported, and intended particle is sandwiched in the groove between the first sub-grip block (051) and the second sub-grip block (052);
Step 4: will be used for the ac voltage signal that particle controls and close; Between the first sub-grip block (051) and the second sub-grip block (052), apply the ac voltage signal that is used for impedance measurement then; Make electric signal pass through tested single-particle; The frequency in conditioning signal source in predetermined scope, and gather the amplitude, phase information of the sine voltage signal on the first sub-grip block (051) and the second sub-grip block (052) in real time and then through calculating the resistance value of tested particle by signal analyzer.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101871974B (en) * | 2010-06-18 | 2013-01-09 | 华南理工大学 | Impedance spectroscopy measurement method |
| TW201816380A (en) * | 2016-10-27 | 2018-05-01 | 統創科技股份有限公司 | Light induced dielectrophoresis (LIDEP) device capable of performing a sorting process on a liquid comprising plural first micro-particles and plural second micro-particles |
| KR20200005466A (en) * | 2018-07-06 | 2020-01-15 | 주식회사 엘지화학 | Surface area measurement method of conductive materials |
| CN109821582B (en) * | 2019-03-13 | 2021-12-03 | 京东方科技集团股份有限公司 | Particle capture structure, particle capture chip and particle capture method |
| CN111890249B (en) * | 2020-07-10 | 2021-11-16 | 东南大学 | Chip clamping and fixing and chip parallelism measuring structure for ultrasonic bonding |
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| CN101344518A (en) * | 2008-08-15 | 2009-01-14 | 东南大学 | Multi-mode set integration dielectric characterization apparatus and method of micro-nano biological particle |
| CN201532352U (en) * | 2009-11-06 | 2010-07-21 | 东南大学 | Micro-nano single particle impedance spectrum measuring chip |
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Patent Citations (2)
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|---|---|---|---|---|
| CN101344518A (en) * | 2008-08-15 | 2009-01-14 | 东南大学 | Multi-mode set integration dielectric characterization apparatus and method of micro-nano biological particle |
| CN201532352U (en) * | 2009-11-06 | 2010-07-21 | 东南大学 | Micro-nano single particle impedance spectrum measuring chip |
Non-Patent Citations (2)
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| H.Morgan et al..High speed simultaneous single particle impedance and fluorescence analysis on a chip.《current applied physics》.2005,第6卷367-370. * |
| 倪中华等.基于光诱导介电泳的微纳米生物粒子操纵平台关键技术.《中国科学 E辑:技术科学》.2009,第39卷(第10期),1635-1642. * |
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