CN115895864A - Micro-fluidic chip detection system based on planar electrode - Google Patents
Micro-fluidic chip detection system based on planar electrode Download PDFInfo
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- CN115895864A CN115895864A CN202211525198.6A CN202211525198A CN115895864A CN 115895864 A CN115895864 A CN 115895864A CN 202211525198 A CN202211525198 A CN 202211525198A CN 115895864 A CN115895864 A CN 115895864A
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- 238000001514 detection method Methods 0.000 title abstract description 11
- 239000000758 substrate Substances 0.000 claims abstract description 16
- 230000002779 inactivation Effects 0.000 claims abstract description 15
- 239000004205 dimethyl polysiloxane Substances 0.000 claims abstract description 11
- 235000013870 dimethyl polysiloxane Nutrition 0.000 claims abstract description 11
- CXQXSVUQTKDNFP-UHFFFAOYSA-N octamethyltrisiloxane Chemical compound C[Si](C)(C)O[Si](C)(C)O[Si](C)(C)C CXQXSVUQTKDNFP-UHFFFAOYSA-N 0.000 claims abstract description 11
- 238000004987 plasma desorption mass spectroscopy Methods 0.000 claims abstract description 11
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 claims abstract description 11
- 239000011521 glass Substances 0.000 claims abstract description 10
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims abstract description 9
- 239000010931 gold Substances 0.000 claims abstract description 9
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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Abstract
The invention relates to the field of microfluidic chips, and discloses a microfluidic chip detection system based on a planar electrode, which comprises a microfluidic chip, an addressing inactivation controller, a pressure controller and a control unit, wherein the microfluidic chip comprises a planar interdigital electrode and a PDMS chamber layer, the PDMS chamber layer comprises a glass substrate and a patterned gold electrode, a channel structure is arranged on the glass substrate, the channel structure comprises two sample inlets and a plurality of capture chambers communicated with the sample inlets, and one sample inlet is communicated with the pressure controller. The invention can realize the single cell accommodation of the capture chamber by designing and optimizing the channel on the chip and the capture chamber mechanism, firstly screens the required target cell area by using a computer program and an addressing fire extinguisher, and sends a logic instruction to load pulse electric signals which are enough to electrocute cells on the control electrodes corresponding to the rest cell areas, and after the inactivation operation is finished, all cells are collected, and the obtained sample only has active target cells.
Description
Technical Field
The invention relates to the field of microfluidic chips, in particular to a microfluidic chip detection system based on a planar electrode.
Background
The microfluidic chip technology integrates basic operation units of sample preparation, reaction, separation, detection and the like in biological, chemical and medical analysis processes into a micron-scale chip, and automatically completes the whole analysis process. Due to its great potential in the fields of biology, chemistry, medicine and the like, the method has been developed into a new research field crossing the disciplines of biology, chemistry, medicine, fluid, electronics, materials, machinery and the like. The micro-fluidic chip analysis takes a chip as an operation platform, simultaneously takes analytical chemistry as a basis, takes a micro-electromechanical processing technology as a support, takes a micro-pipeline network as a structural characteristic, takes life science as a main application object at present, and is the key point of the development of the field of the current micro total analysis system.
The cell is a basic unit for forming life, and the research on the structure and function of the cell has great significance for the exploration of life rules and the diagnosis and treatment of diseases. At present, with the intensive research on cell specificity, single cell/single class cell analysis and screening also become a hot spot in the field of life science research. Therefore, the inventor aims to develop a microfluidic chip detection system, and the non-target cells are killed reversely so as to realize the screening and acquisition of specific target cells.
Disclosure of Invention
The invention aims to provide a micro-fluidic chip detection system based on a planar electrode, so as to realize screening and acquisition of target cells through killing of non-target cells.
In order to achieve the purpose, the invention adopts the following technical scheme: the utility model provides a micro-fluidic chip detecting system based on plane electrode, includes micro-fluidic chip, addressing formula inactivation control ware, pressure controller and the control unit, and micro-fluidic chip includes plane interdigital electrode and PDMS chamber layer, and PDMS chamber layer includes glass substrate and patterning gold electrode, is provided with the channel structure on the glass substrate, the channel structure includes two introduction ports and a plurality of capture cavity with the introduction port intercommunication, and the wide 5 mu m of earial drainage department of capture cavity, the wide 16 mu m of cup-shaped structure of capture cavity, one of them introduction port and pressure controller intercommunication.
The principle and the advantages of the scheme are as follows: in practical application, microfluidics is a technology for processing or manipulating fluid, and the current cell manipulation methods based on microfluidics are mainly divided into an active type and a passive type, wherein the active type refers to the fact that external force generated by active control is utilized to achieve manipulation of fluid and cells, common methods include valve control, electric control, magnetic control, light control and sound control, and the method has the advantages of good controllability, high accuracy and the like. The passive mode is to use the interactive coupling action of the structure and the fluid, the cell and the fluid and the cell and the structural part, namely the fluid mechanics principle to realize the accurate control of the movement of the fluid and the cell in the flow channel, and has the characteristics of simple system, high sample processing flux and the like. In the previous research, the microfluidic chip device uses dielectrophoresis to push the cells trapped in the chamber out of the chamber, but the damage of the cells under the electrical stimulation is not easy to control, and in the previous research and development process, it is found that the cells in the buffer solution such as PM, PBS, etc. are easily electrocuted in situ, so the reverse thinking is to think that the whole row of the unwanted cells is inactivated, and the wanted target cells are left. Therefore, the technical scheme is based on sample hydrodynamics, only single cell accommodation can be realized in the capture cavity through the design and optimization of the channel on the chip and the capture cavity mechanism, a required target cell area is firstly screened by utilizing a computer program and an addressing fire extinguisher, a pulse electric signal which is enough to electrocute cells is loaded to control electrodes corresponding to other cell areas through a logic command issued by a computer so as to kill non-target cells, and in the process, voltage parameters need to be gradually increased so as to avoid overhigh voltage; and the power-on needs to be performed sequentially and needs to be performed region by region, so that the situation that the power of the region needing to be subjected to power-on inactivation is added at one time is avoided, and the influence of an overhigh joule heat phenomenon in the cavity on the survival cells is prevented. Furthermore, the time required to control the power-up is not likely to be too long. Normally, the voltage is controlled to be about 15V, and the electric signal is a pulse signal: the pulse width is 100 mus, each pulse is 1s apart, and a single electrical signal covers 5 pulses. After the inactivation operation is completed, all cells are collected, and only active target cells exist in the obtained sample. The technical scheme avoids the electrical stimulation damage of cells in the prior art from the source, not only can accurately kill non-target cells, but also can synchronously and efficiently sort the target cells and the non-target cells.
Preferably, as an improvement, the trapping chambers are arranged in an array, and the column pitch and the row pitch are both 220 μm.
In the technical scheme, the plurality of capture chamber arrays are arranged, so that the detection flux can be improved; in addition, the distance between the capture chambers needs to be comprehensively considered based on the overall size of the chip and the observation convenience, the distance is too large, the overall size of the chip can be enlarged, the observation is inconvenient, the distance is too small, and the electric interaction between the adjacent chambers can be large.
Preferably, as a modification, the planar interdigital electrode comprises a base layer and a patterned gold electrode disposed on the base layer, and the patterned gold electrode comprises a plurality of common electrodes and a plurality of independent control electrodes.
In the technical scheme, the electric field subsections corresponding to the capture chambers have key influence on killing and sorting of target cells, and the quantity of the throughput electrodes is set to be multiple, so that high capture efficiency can be guaranteed on one hand, and the high flux of detection can be met on the other hand.
Preferably, as a modification, the substrate layer is a glass substrate.
In the technical scheme, the glass substrate has stable property and is not influenced by an oxidizing agent, a reducing agent and other impurities.
Preferably, as a modification, the width of each of the common electrode and the independent control electrode is 100 μm.
In the technical scheme, the electrode width is firstly set based on the chamber distance, on the basis, the wider the electrode is, the smaller the resistance is, and the smaller the attenuation degree received by the electric signal transmission is when the power generation signal is generated.
Preferably, as a modification, the number of the trapping chambers is 128, and the arrangement of the 128 trapping chambers is: each column has 16 rows and 8 columns, the row direction is divided into 4 groups, each group has 2 rows, the row spacing in the group is 220 μm, and the row spacing between the groups is 2120 μm.
Among this technical scheme, the distribution of catching the cavity is based on chip mechanism on the one hand, and on the other hand can guarantee high-efficient killing and avoid influencing each other between the adjacent cavity through the optimization to inter block line space and inter block line space.
Preferably, as an improvement, the control unit is a computer operating platform.
In the technical scheme, the computer operating platform is adopted to screen the target cell area and issue the logic instruction to control the electrode of the corresponding area to load the pulse electric signal which is enough to electrocute the cell, so that the operation is convenient and the accuracy is high.
Drawings
Fig. 1 is a schematic diagram of a microfluidic chip detection system based on a planar electrode according to an embodiment of the present invention.
Fig. 2 is a schematic structural diagram of the microfluidic chip.
FIG. 3 is a schematic view of a channel structure.
FIG. 4 is a schematic diagram of the connection between the capture chamber and the sample channel.
FIG. 5 is a micrograph showing the results of the validation of the inactivation mode in the example of the present invention.
Detailed Description
The following is a detailed description of the embodiments, but the embodiments of the present invention are not limited thereto. Unless otherwise specified, the technical means used in the following embodiments are conventional means well known to those skilled in the art; the experimental methods used are all conventional methods; the materials, reagents and the like used are all commercially available.
Reference numerals in the drawings of the specification include: the device comprises a micro-fluidic chip 1, an addressing inactivation controller 2, a pressure controller 3, a computer operating platform 4, a planar interdigital electrode 5, a PDMS chamber layer 6, a channel structure 7, a common electrode 8, an independent control electrode 9, a capture chamber 10, a sample inlet 11 and a sample injection channel 12.
Example 1
As shown in fig. 1-4, a microfluidic chip detection system based on planar electrodes includes a microfluidic chip 1, an addressable inactivation controller 2, a pressure controller 3, and a computer operation platform 4. The technical scheme mainly comprises the steps of improving and designing the structure of the microfluidic chip 1, wherein the addressing type inactivation controller 2 and the computer operation platform 3 are both used and connected in the prior art.
The microfluidic chip 1 comprises a planar interdigital electrode 5 and a PDMS chamber layer 6, wherein a channel structure 7 is arranged in the PDMS chamber layer 6.
The structure of the planar interdigital electrode 5 is shown in fig. 4, and comprises a glass substrate and a patterned gold electrode, wherein the patterned gold electrode comprises four common electrodes 8 with the width of 100 micrometers and eight independent control electrodes 9 with the width of 100 micrometers; and the common electrodes 8 are independent of each other.
The channel structure 7 includes 128 trapping chambers 10 disposed in the PDMS chamber layer, and the 128 trapping chambers 10 are arranged in the following manner: 16 per column and 8 per row; the column pitch was 220 μm, the row direction was divided into 4 groups of 2 rows, the row pitch within the group was 220 μm, and the row pitch between the groups was 2120 μm. Be provided with introduction port 11 on the PDMS, introduction port 11 is provided with two, all communicate sample channel 12 between two introduction port 11 and the capture chamber 10, and one of them introduction port 11 is kept away from the one end of capturing chamber 10 and is used for the appearance of sample, and another introduction port 11 communicates with pressure controller 3, so, accessible pressure controller 3 makes the sample get into sample channel 12 under the state of negative pressure to under the control of fluid power, make the cell fixed in capture chamber 10.
When the technical scheme is actually used, a sample to be sorted enters the sample inlet channel 12 along the sample inlet, the pressure controller 3 is started at the same time, the sample enters the capture chamber 10 along the sample inlet channel 12 under the control of negative pressure, and cells are fixed in the capture chamber 10 under the control of fluid force. The computer program and the addressing type inactivation controller 2 are utilized to screen the required target cell area firstly, the pulse electric signals which are enough to electrocute the cells are loaded to the control electrodes corresponding to the other cell areas through the logic instructions issued by the computer, after the inactivation operation is finished, all the cells are collected, and the obtained sample only has the active target cells.
Experimental example-verification experiment of inactivation mode
The experiment utilizes a single control electrode (6 pairs) to carry out selective killing experiments on 6 chambers. The experimental method is as follows:
1. aligning the micro-fluidic chip and the microelectrode under a microscope;
2. centrifuging the cells, suspending the cells in a PM buffer solution, introducing the cell suspension into a microfluidic chip, and capturing the cells in a chamber under the action of a fluid force;
3. target cell sites (site 6 is the site of the tap, corresponding to the control site on the far right in FIG. 5) were selected and the voltage (15V) was applied to the electrode pairs corresponding to the remaining sites. The voltage signal is a pulse signal, the pulse width is 100 mus, the pulse period is 1s, and the number of pulses is 5. The experiment was repeated, and when the signal amplitude was increased to 15V, cells were inactivated at each site. Waiting for about 3 minutes after each power-on, avoiding continuous power-on because the cells are not observed to be electrocuted, and avoiding some reaction time of the cells under the action of electricity.
4. After the inactivation operation is completed, trypan blue solution can be introduced to stain the cells, dead cells can be stained into blue, and living cells can not be stained, so that the feasibility of the inactivation operation is proved.
The results of the experiment are shown in FIG. 5, and show that only 6-th site survived after trypan blue staining, and the rest were electrocuted.
The above description is only an example of the present invention, and the general knowledge of the known specific technical solutions and/or characteristics and the like in the solutions is not described herein too much. It should be noted that, for those skilled in the art, without departing from the technical solution of the present invention, several variations and modifications can be made, which should also be regarded as the protection scope of the present invention, and these will not affect the effect of the implementation of the present invention and the practicability of the patent. The scope of the claims of the present application shall be determined by the contents of the claims, and the description of the embodiments and the like in the specification shall be used to explain the contents of the claims.
Claims (7)
1. The utility model provides a micro-fluidic chip detecting system based on plane electrode which characterized in that: including micro-fluidic chip, addressing formula inactivation control ware, pressure controller and the control unit, micro-fluidic chip includes plane interdigital electrode and PDMS chamber layer, and PDMS chamber layer includes glass substrate and patterning gold electrode, is provided with the channel structure on the glass substrate, the channel structure includes two introduction port and a plurality of and the capture cavity of introduction port intercommunication, the wide 5 mu m of bleeder department of capture cavity, the wide 16 mu m of cup-shaped structure of capture cavity, one of them introduction port and pressure controller intercommunication.
2. The system according to claim 1, wherein the micro-fluidic chip comprises a substrate, a micro-fluidic chip, and a planar electrode, wherein the micro-fluidic chip comprises: the capture chambers are arranged in an array, and the row spacing and the column spacing are both 220 micrometers.
3. The system of claim 2, wherein the micro-fluidic chip comprises a planar electrode, and the system further comprises: the planar interdigital electrode comprises a substrate layer and a patterned gold electrode arranged on the substrate layer, wherein the patterned gold electrode comprises a plurality of common electrodes and a plurality of independent control electrodes.
4. The system according to claim 3, wherein the micro-fluidic chip comprises a substrate, a micro-fluidic chip, and a micro-fluidic chip, wherein the micro-fluidic chip comprises: the substrate layer is a glass substrate.
5. The system according to claim 4, wherein the micro-fluidic chip comprises a planar electrode, and the system further comprises: the width of the common electrode and the width of the independent control electrode are both 100 mu m.
6. The system of claim 5, wherein the micro-fluidic chip comprises a planar electrode, and the system further comprises: the number of the capture chambers is 128, and the arrangement of the 128 capture chambers is as follows: each column has 16 rows and 8 columns, the row direction is divided into 4 groups, each group has 2 rows, the row spacing in the group is 220 μm, and the row spacing between the groups is 2120 μm.
7. The system according to claim 6, wherein the micro-fluidic chip comprises a planar electrode, and the system further comprises: the control unit is a computer operating platform.
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| CN202211525198.6A CN115895864A (en) | 2022-11-30 | 2022-11-30 | Micro-fluidic chip detection system based on planar electrode |
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