CN113866074A - Impedance method's leucocyte classification count micro-fluidic chip based on position compensation - Google Patents
Impedance method's leucocyte classification count micro-fluidic chip based on position compensation Download PDFInfo
- Publication number
- CN113866074A CN113866074A CN202111137092.4A CN202111137092A CN113866074A CN 113866074 A CN113866074 A CN 113866074A CN 202111137092 A CN202111137092 A CN 202111137092A CN 113866074 A CN113866074 A CN 113866074A
- Authority
- CN
- China
- Prior art keywords
- electrodes
- differential
- electrode
- impedance method
- pulse
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000000034 method Methods 0.000 title claims abstract description 34
- 210000000265 leukocyte Anatomy 0.000 claims abstract description 21
- 239000012530 fluid Substances 0.000 claims abstract description 12
- 238000012545 processing Methods 0.000 claims abstract description 6
- 210000004027 cell Anatomy 0.000 claims description 46
- 238000010586 diagram Methods 0.000 claims description 7
- 230000009471 action Effects 0.000 claims description 6
- 230000033001 locomotion Effects 0.000 claims description 6
- 238000005070 sampling Methods 0.000 claims description 6
- 239000000243 solution Substances 0.000 claims description 6
- 239000008151 electrolyte solution Substances 0.000 claims description 5
- 230000008859 change Effects 0.000 claims description 4
- 239000003085 diluting agent Substances 0.000 claims description 4
- 230000003321 amplification Effects 0.000 claims description 3
- 230000001066 destructive effect Effects 0.000 claims description 3
- 238000009792 diffusion process Methods 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 3
- 238000001514 detection method Methods 0.000 abstract description 12
- 230000004907 flux Effects 0.000 abstract description 3
- 230000035945 sensitivity Effects 0.000 abstract description 3
- 230000002950 deficient Effects 0.000 abstract description 2
- 238000003745 diagnosis Methods 0.000 abstract description 2
- 238000009826 distribution Methods 0.000 description 11
- 210000004369 blood Anatomy 0.000 description 5
- 239000008280 blood Substances 0.000 description 5
- 230000008569 process Effects 0.000 description 3
- 230000002902 bimodal effect Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000000386 microscopy Methods 0.000 description 2
- 238000000790 scattering method Methods 0.000 description 2
- 206010018910 Haemolysis Diseases 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 238000000149 argon plasma sintering Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 210000000601 blood cell Anatomy 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 238000012937 correction Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 230000005684 electric field Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 238000001917 fluorescence detection Methods 0.000 description 1
- 230000008588 hemolysis Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume, or surface-area of porous materials
- G01N15/10—Investigating individual particles
-
- G01N2015/1024—
Abstract
The invention relates to the technical field of fluid chips, in particular to a leukocyte differential counting microfluidic chip based on an electrical impedance method of position compensation. This leucocyte differential count micro-fluidic chip based on electrical impedance method of position compensation, the double differential structure electrode has been integrated, have high flux, the micro-sample, exempt from the label, characteristics such as high sensitivity, can count and classify the leucocyte fast accurately, and the size is little, the integrated level is high, the light-dutyization of possible product after the processing is conveniently carried, easy operation weak point consuming time need not professional skill or extra medical instrument, even be the poverty area that remote medical treatment condition is deficient also can realize the timely detection in the daily diagnosis, the error of getting rid of artifical count reduces personnel and infects the risk.
Description
Technical Field
The invention relates to the technical field of fluid chips, in particular to a leukocyte differential counting microfluidic chip based on an electrical impedance method of position compensation.
Background
The leucocyte plays an important role in the blood cell of the human body, can generate an antibody to resist the invasion of external viruses and cure the damage of the function of the human body, and the microfluidic chip is a system which controls a target object with the scale of several micrometers or even nanometer level in a channel with the scale of tens of micrometers to hundreds of micrometers so as to ensure the target object to move regularly and orderly.
The current counting and classifying method of white blood cells mainly comprises a manual microscopy, a laser scattering method and an electrical impedance method, wherein the manual microscopy is that a professional carries out hemolysis dilution on a certain amount of blood, dissolves red blood cells in the blood, discharges interference on the white blood cells, drops the dissolved liquid into a counting pool of a special cell counting plate, carries out manual counting on the cells in a specific area under a microscope, and finally converts the content of the white blood cells in each liter of blood; the laser scattering method is a principle of light transmission to realize cell classification, different objects are irradiated with different light scattering characteristics, light is transmitted in a straight line in uniform cut-off, however, when the device passes through a non-uniform medium such as cell suspension liquid, a part of light can be transmitted and absorbed, the rest part of light can be scattered from the original direction, the angle and distribution of the scattered light contain characteristic information such as the size, the refractive index, the symmetry and the like of cells, and therefore, the laser scattering can be utilized to determine the size and the shape of the cells to realize classification and counting.
However, the manual microscope method needs trained professional counting personnel to complete, the technical requirement is high, time is long and large inevitable errors of manual counting can occur due to manual completion, meanwhile, the personnel are exposed for a long time, and the risk of virus infection exists when the sample is contacted, so that the manual microscope method is not suitable for large-scale popularization and the application scene is limited; the laser detector detects accumulated scattered light within a certain angle range, and for cell populations with broadband or bimodal size distribution, the situation that the accumulated scattered light is generated, but the cell morphology size, the refractive index and the like are different occurs, so that counting errors are generated, and useful information for cell classification is lost, and meanwhile, devices which are unavailable for laser scattering counting are a laser source and a detector, which bring unavoidable defects of high cost, size incapability of miniaturization, weight increase and the like to products.
Disclosure of Invention
The invention aims to provide a leukocyte differential counting microfluidic chip based on an electrical impedance method of position compensation, so as to solve the problems in the background technology.
In order to achieve the purpose, the invention provides the following technical scheme: the leukocyte differential counting microfluidic chip based on the position compensation electrical impedance method comprises a microfluidic chip, wherein the microfluidic chip comprises a fluid channel, electrolyte solution is filled in the fluid channel, seven electrodes are symmetrically distributed at the bottom of the fluid channel at equal intervals, and the microfluidic chip adopts a double differential structure.
Preferably, among the seven electrodes, the 1 st and 7 th electrodes are signal input electrodes, the 2 nd and 6 th electrodes are floating electrodes, the 3 rd and 5 th electrodes are reference ground electrodes, the 4 th electrode, i.e., the center electrode, is also a signal input electrode, and the 4 th electrode is connected with an alternating current signal having a phase opposite to that of the 1 st and 7 th electrodes.
Preferably, the leukocyte classification method based on the position compensated electrical impedance method comprises the following steps:
s1: applying alternating voltage with specific amplitude and frequency to the central electrode, and applying voltages with opposite phases to the electrodes (No. 1 and No. 7) at the two ends;
s2: adding the treated leukocyte diluent into a sample reservoir, opening a micro pump to generate negative pressure to pump a cell solution into a channel, enabling the cells to move forwards along the channel along the direction of an arrow in the figure under the action of pressure and diffusion movement, and enabling the cells to instantaneously cause pulse change when moving through different electrodes to generate current signals;
s3: sending the two paths of pulse current signals after differential output to an IV converter for destructive filtering, and then carrying out differential amplification to remove internal noise current signals of a channel and the IV converter;
s4: sampling the output pulse signal at a certain sampling rate, and transmitting the digitized signal to a computer for further signal processing to finally generate a pulse schematic diagram;
s5: when the cell passes through the ground reference electrode and the central electrode, an inverted pulse is generated, the amplitude of the inverted peak is much larger than that of the previous floating electrode, and therefore the cell can be counted and classified in the area according to the pulse amplitude.
Compared with the prior art, the invention has the beneficial effects that:
1. the impedance method based on position compensation leukocyte differential counting microfluidic chip integrates the electrodes with double differential structures, has the characteristics of high flux, micro-samples, no label, high sensitivity and the like, and can be used for rapidly and accurately counting and classifying leukocytes.
2. The leucocyte differential count micro-fluidic chip based on the electrical impedance method of position compensation is small in size and high in integration level, a processed product can be light and convenient to carry, errors of manual counting are eliminated, and personnel infection risks are reduced.
3. The impedance method based on the position compensation leukocyte differential counting microfluidic chip is simple to operate, consumes short time, does not need professional skills or additional medical instruments, and can realize timely detection in daily diagnosis even in remote poor areas with deficient medical conditions.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings needed to be used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without inventive labor.
FIG. 1 is a schematic structural diagram of a microfluidic chip according to the present invention;
FIG. 2 is a schematic diagram of a double differential structure according to the present invention;
FIG. 3 is a schematic diagram of the detection signal of the present invention;
FIG. 4 is a graph of the original detection signal distribution of the present invention;
FIG. 5 is a calibrated detection signal distribution diagram according to the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In the present invention, unless otherwise expressly stated or limited, the terms "mounted," "connected," "secured," and the like are to be construed broadly and can, for example, be fixedly connected, detachably connected, or integrally formed; can be mechanically or electrically connected; either directly or indirectly through intervening media, either internally or in any other relationship. The specific meanings of the above terms in the present invention can be understood by those skilled in the art according to specific situations.
Referring to fig. 1-5, the present invention provides a technical solution: the leucocyte classification counting micro-fluidic chip based on the position compensation electrical impedance method comprises a micro-fluidic chip, wherein the micro-fluidic chip comprises a fluid channel, electrolyte solution is filled in the fluid channel, the electrolyte solution has good conductivity, a current loop is formed in the channel, seven electrodes are symmetrically distributed at the bottom of the fluid channel at equal intervals, of the seven electrodes, the No. 1 and No. 7 are signal input electrodes and are used for being connected with an external alternating current power supply, the No. 2 and No. 6 electrodes are floating electrodes used for cell calibration, the No. 3 and No. 5 electrodes are reference ground electrodes, a central electrode is also a signal input electrode and is connected with alternating current signals with opposite phases of the No. 1 and No. 7 electrodes, and the micro-fluidic chip adopts a double differential structure.
The first differential stage in the double differential structure in the chip is: respectively leading out signals of the No. 1 pair of reference ground electrodes and signals of the central electrode and the reference ground electrodes, and passing through a differential amplifier; similarly, signals of the No. 7 chip right side and the central electrode relative to the reference ground electrode are led out and output through a differential amplifier for subsequent signal processing; a second difference stage: the two paths of ground differential output signals are sent to an IV converter and then pass through a differential amplifier; as shown in fig. 2: alternating voltages with opposite phases are applied to the No. 1 and No. 7 electrodes and the central electrode, and the channels are filled with electrolyte solution, so that weak electric signals can be detected from the electrodes on the lower substrate; when cells flow through the channel, the balance condition in the channel can be broken, different pulse signals are formed between the central electrode and the two side electrodes, the generated pulse signals are enhanced and output after passing through the differential circuit, the detection and counting of the cells are realized, and meanwhile, the pulse signals generated when different cells pass through the channel are different in size, so that the classification of the cells is realized.
The white blood cell classification method based on the position compensation electrical impedance method comprises the following steps:
s1: applying alternating voltage with specific amplitude and frequency to the central electrode, and applying voltages with opposite phases to the electrodes (No. 1 and No. 7) at the two ends;
s2: adding the well processed leukocyte diluent into a sample reservoir, opening a micro pump to generate negative pressure to pump a cell solution into a channel, enabling cells to move forward along the channel along the direction of an arrow in the figure under the action of pressure and diffusion movement, and enabling the cells to instantaneously cause pulse change when moving through different electrodes to generate current signals, wherein the change becomes more obvious due to the adoption of a differential structure;
s3: for subsequent signal processing, the two paths of pulse current signals after differential output are sent to an IV converter for destructive filtering, and then internal noise current signals of a channel and the IV converter are removed through differential amplification;
s4: sampling the output pulse signal at a certain sampling rate, transmitting the digitized signal to a computer for further signal processing, and finally generating a pulse schematic diagram, as shown in fig. 3, the reason for generating the double-peak pulse is that the floating electrode causes the non-uniformity of electric field distribution, and simultaneously, different positions of the movement of the cell passing through the channel can influence the height of a trough between the double peaks, wherein the movement positions refer to the movement of the cell approaching the top, the middle and the bottom of the channel;
s5: when the cell passes through the ground reference electrode and the central electrode, an inverted pulse is generated, the amplitude of the inverted peak is much larger than that of the previous floating electrode, and therefore the cell can be counted and classified in the area according to the pulse amplitude.
The position of a cell moving in a fluid channel influences the magnitude of a pulse signal, the amplitude of current generated when the same cell passes through the channel at different vertical heights is different, meanwhile, a cell with a larger size often flows at a higher position, and a cell with a smaller size often moves at a lower position, so that the magnitude of the pulse generated near a central electrode alone is not enough to accurately measure and classify the cell, and therefore, the error caused by the moving position of the cell needs to be corrected to ensure the accuracy of cell sample measurement, and the floating electrode is used for the purpose.
The impedance method based on position compensation can realize rapid counting and classification of white blood cells only by trace samples, is different from the conventional detection of hospital blood in that a large amount of blood samples need to be collected for detection, is different from the laser scattering detection that an expensive laser light source is needed, and is different from the fluorescence detection that the samples need to be subjected to labeling treatment.
The two-sided floating electrode area is defined as PF:
where P is the peak and trough of a doublet, respectively, and PF is a defined number between 0 and 1, used to correct the cells.
The middle area of fig. 3 is defined as the area of the original signal size RES, and the amplitude of the signal generated in the area is defined as a:
where G is a geometric parameter whose value is determined by the channel dimensions.
Taking PF as reference to normalize the original RES, and defining the processed result as NES:
where the RES parameters are as described above, BeadSize is the standard cell size used for calibration.
As shown in fig. 4, the upper half of the graph is a histogram of counts measured when a leukocyte diluent of a certain standard size passes through the microfluidic chip, the lower half of the graph is a distribution of a scattergram in which PF corresponding to the standard cell is taken as a vertical axis and RES is taken as a horizontal axis, the histogram is not normalized, the number of times of occurrence of an original signal RES of the cell is counted, the reason of occurrence of different distributions is a difference in vertical positions at which the cell of the certain type moves in a channel, it can be seen from the scattergram that the particle distribution approximately presents a straight line, and a two-dimensional NES ═ c is established for the distribution1×PF+c2The model, using a linear fitting algorithm, solves for the sum of the correction parameters, since the linear fitting parameter c1、c2The method is solved on the basis of the normalized dimension NES, so that the method is also suitable for cells with other dimensions which agree with detection environment detection, and can be used for the subsequent dimension detection of any cell only by once calibration.
Dividing the original signal size RES by the calibration parameter to obtain a calibrated size expression CES:
fig. 5 shows the cell size after calibration, and it can be seen that the distribution graph of the original bimodal shape after calibration becomes approximately standard normal unimodal distribution, and the mean value is approximately the actual size of the cell.
Generally speaking, the impedance method based on position compensation leukocyte differential counting microfluidic chip integrates the electrodes with double differential structures, has the characteristics of high flux, micro samples, label-free, high sensitivity and the like, and can be used for rapidly and accurately counting and classifying leukocytes.
It is noted that, herein, relational terms such as first and second, and the like may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. The term "comprising", without further limitation, means that the element so defined is not excluded from the group consisting of additional identical elements in the process, method, article, or apparatus that comprises the element.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (3)
1. The leucocyte differential count micro-fluidic chip based on the electrical impedance method of position compensation comprises a micro-fluidic chip and is characterized in that: the microfluidic chip comprises a fluid channel, electrolyte solution is filled in the fluid channel, seven electrodes are symmetrically distributed at the bottom of the fluid channel at equal intervals, and the microfluidic chip adopts a double-differential structure.
2. The leukocyte differential counting microfluidic chip based on the electrical impedance method of position compensation of claim 1, wherein: of the seven electrodes, the No. 1 and No. 7 are signal input electrodes, the No. 2 and No. 6 electrodes are floating electrodes, the No. 3 and No. 5 electrodes are reference ground electrodes, the No. 4 electrode, namely the central electrode, is also a signal input electrode, and the No. 4 electrode is connected with an alternating current signal with opposite phase to the No. 1 and No. 7 electrodes.
3. The leukocyte differential counting microfluidic chip based on the electrical impedance method of position compensation of claim 1, wherein: the leucocyte classification method based on the electrical impedance method comprises the following steps:
s1: applying alternating voltage with specific amplitude and frequency to the central electrode, and applying voltages with opposite phases to the electrodes (No. 1 and No. 7) at the two ends;
s2: adding the treated leukocyte diluent into a sample reservoir, opening a micro pump to generate negative pressure to pump a cell solution into a channel, enabling the cells to move forwards along the channel along the direction of an arrow in the figure under the action of pressure and diffusion movement, and enabling the cells to instantaneously cause pulse change when moving through different electrodes to generate current signals;
s3: sending the two paths of pulse current signals after differential output to an IV converter for destructive filtering, and then carrying out differential amplification to remove internal noise current signals of a channel and the IV converter;
s4: sampling the output pulse signal at a certain sampling rate, and transmitting the digitized signal to a computer for further signal processing to finally generate a pulse schematic diagram;
s5: when the cell passes through the ground reference electrode and the central electrode, an inverted pulse is generated, the amplitude of the inverted peak is much larger than that of the previous floating electrode, and therefore the cell can be counted and classified in the area according to the pulse amplitude.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111137092.4A CN113866074A (en) | 2021-09-27 | 2021-09-27 | Impedance method's leucocyte classification count micro-fluidic chip based on position compensation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111137092.4A CN113866074A (en) | 2021-09-27 | 2021-09-27 | Impedance method's leucocyte classification count micro-fluidic chip based on position compensation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113866074A true CN113866074A (en) | 2021-12-31 |
Family
ID=78991285
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111137092.4A Pending CN113866074A (en) | 2021-09-27 | 2021-09-27 | Impedance method's leucocyte classification count micro-fluidic chip based on position compensation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113866074A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116735461A (en) * | 2023-08-10 | 2023-09-12 | 成都云芯医联科技有限公司 | Three categorised optical detection systems of leucocyte |
-
2021
- 2021-09-27 CN CN202111137092.4A patent/CN113866074A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116735461A (en) * | 2023-08-10 | 2023-09-12 | 成都云芯医联科技有限公司 | Three categorised optical detection systems of leucocyte |
| CN116735461B (en) * | 2023-08-10 | 2023-11-10 | 成都云芯医联科技有限公司 | Three categorised optical detection systems of leucocyte |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US10670508B2 (en) | Microfluidic device for selection of semen | |
| US20220118444A1 (en) | Microchip and particulate analyzing device | |
| US9074978B2 (en) | Optical space-time coding technique in microfluidic devices | |
| EP3220130B1 (en) | High accuracy 5-part differential with digital holographic microscopy and untouched leukocytes from peripheral blood | |
| CN103323383B (en) | Particle counting system of micro-fluidic chip based on electric resistance technology | |
| EP2980558B1 (en) | Measurement device and measurement method | |
| CN106644900B (en) | Impedance pulse particle counting device based on non-uniform electric field and counting method thereof | |
| US20100020308A1 (en) | Transducer Module | |
| CN102239400A (en) | Sheath-fluid-free flow cytometry method and device | |
| CN203337513U (en) | Micro-fluidic chip particle counting system based on electrical impedance technology | |
| CN113866074A (en) | Impedance method's leucocyte classification count micro-fluidic chip based on position compensation | |
| CN111359687B (en) | Micro-fluidic chip and blood cell analysis device based on electricity and fluorescence signals | |
| Lee et al. | A CMOS impedance cytometer for 3D flowing single-cell real-time analysis with ΔΣ error correction | |
| CN113567326A (en) | High-throughput real-time single-cell electrical intrinsic parameter measurement system and method | |
| CN204405503U (en) | A kind of portable quick particle detection | |
| CN207964570U (en) | A kind of adjustable grain count device of accuracy of detection | |
| CN101025414A (en) | Laser blood cell counter | |
| WO2023218481A1 (en) | A microfluidic impedance cytometry apparatus | |
| CN203249851U (en) | Cell scattering light multi-position detection cytometer | |
| CN114345428B (en) | Micro-fluidic chip for selecting single cells and detection method | |
| CN108279202A (en) | A kind of adjustable grain count device and method of accuracy of detection | |
| CN103278440A (en) | Cell scattered light multi-position detecting method as well as cell analyzer | |
| CN108226013B (en) | Device and method for improving particle detection precision and flux by electric field focusing | |
| CN208155808U (en) | A kind of device improving particle detections precision and flux using static focusing | |
| CN106290081A (en) | A kind of method distinguishing particle group and particle analyzer |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |