CN112378827A - Wide particle detection device of size range - Google Patents
Wide particle detection device of size range Download PDFInfo
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- CN112378827A CN112378827A CN202011330230.6A CN202011330230A CN112378827A CN 112378827 A CN112378827 A CN 112378827A CN 202011330230 A CN202011330230 A CN 202011330230A CN 112378827 A CN112378827 A CN 112378827A
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- 238000001514 detection method Methods 0.000 title claims abstract description 182
- 239000002245 particle Substances 0.000 title claims abstract description 157
- 230000003321 amplification Effects 0.000 claims abstract description 20
- 239000011521 glass Substances 0.000 claims abstract description 20
- 238000003199 nucleic acid amplification method Methods 0.000 claims abstract description 20
- 239000000758 substrate Substances 0.000 claims abstract description 19
- 239000004205 dimethyl polysiloxane Substances 0.000 claims abstract description 14
- 235000013870 dimethyl polysiloxane Nutrition 0.000 claims abstract description 14
- 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 14
- 238000004987 plasma desorption mass spectroscopy Methods 0.000 claims abstract description 14
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 claims abstract description 14
- 238000004458 analytical method Methods 0.000 claims abstract description 9
- 239000000243 solution Substances 0.000 claims description 37
- 238000000520 microinjection Methods 0.000 claims description 35
- 239000012530 fluid Substances 0.000 claims description 33
- 239000007788 liquid Substances 0.000 claims description 10
- 239000007853 buffer solution Substances 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 7
- 238000002347 injection Methods 0.000 claims description 3
- 239000007924 injection Substances 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 238000003825 pressing Methods 0.000 claims description 3
- 239000003153 chemical reaction reagent Substances 0.000 claims description 2
- 238000005516 engineering process Methods 0.000 abstract description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 9
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 6
- 238000010586 diagram Methods 0.000 description 5
- 230000005684 electric field Effects 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 239000008367 deionised water Substances 0.000 description 4
- 229910021641 deionized water Inorganic materials 0.000 description 4
- 238000004140 cleaning Methods 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N glycerol group Chemical group OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 239000004809 Teflon Substances 0.000 description 2
- 229920006362 Teflon® Polymers 0.000 description 2
- -1 flow ratio Substances 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 239000004743 Polypropylene Substances 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 238000006073 displacement reaction Methods 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000011089 mechanical engineering Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 229920001155 polypropylene Polymers 0.000 description 1
- 238000012805 post-processing Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000011897 real-time detection Methods 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000011895 specific detection Methods 0.000 description 1
- 238000004506 ultrasonic cleaning Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
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- 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
- G01N15/06—Investigating concentration of particle suspensions
- G01N15/0656—Investigating concentration of particle suspensions using electric, e.g. electrostatic methods or magnetic methods
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502753—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502761—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules
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- 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
- G01N15/1031—Investigating individual particles by measuring electrical or magnetic effects
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/02—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance
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Abstract
The invention discloses a particle detection device with a wide size range, which belongs to the technical field of particle detection and comprises a detection chip for detecting particles, a signal amplification module for amplifying an electric signal generated after the detection chip detects the particles, and a signal acquisition and analysis module for acquiring the signal amplified by the signal amplification module and counting particles with different sizes respectively; the detection chip is formed by bonding an ITO conductive glass substrate and a PDMS micro-current control chip; the ITO glass substrate is provided with an ITO driving electrode group serving as a driving electrode and an ITO detection electrode group serving as a detection electrode; the ITO driving electrode group is connected with an external power supply through an interface I; based on the impedance pulse sensing detection technology, the accurate detection of particles in a wide size range is realized by utilizing the flow inertial lift force and simultaneously adopting two modes of 'passing through the detection port' and 'not passing through the detection port', and the particles in the range of 0.5-50 mu m can be accurately detected by using the device provided by the invention.
Description
Technical Field
The invention relates to the technical field of particle detection in a micro-channel, in particular to a particle detection device with a wide size range.
Background
In the fields of environmental monitoring, biomedicine, mechanical engineering and the like, the accurate counting of target particles in a sample is of great significance. The detection means commonly used at present mainly takes impedance pulse sensing (RPS) detection technology as the main technology. The technology is that after a direct current or alternating current electric field is applied to two ends of a detection port, when particles flow through the detection port, voltage difference between two ends of the detection port is subjected to sudden change, and the particles can be detected by collecting a sudden change voltage difference signal. Wherein the signal amplitude is related to the particle size and the signal number is consistent with the particle number.
Impedance pulse sensing (RPS) detection techniques take the form of particles passing through a detection port and particles not passing through the detection port. For the form of particles passing through the detection port, if the detection port is too small, the particles are easily blocked by larger-sized particles, and if the detection port is too large, the particles with smaller sizes cannot be detected; for the form that the particles do not pass through the detection port, if the detection port is too small, the identification capability of the particles with larger size is reduced, and if the detection port is too large, the particles with small size are not sensitive. Due to the above contradiction, the existing particle detection technology can only realize accurate detection of particles in a narrow size range. But the various biochemical samples in reality tend to possess a large size distribution range.
When the particle solution is extruded to one side of the channel by using the sheath fluid, the particles with different sizes can generate different degrees of transverse displacement under the action of the flow inertia lifting force, and the particles with smaller sizes are more close to the channel wall, and the particles with larger sizes are more close to the center of the channel.
Disclosure of Invention
The present particle detection technology can only realize the accurate detection of particles with a narrow size range, and for particles with a wide size range, complex post-processing is often required to be carried out on detection signals, and real-time online observation cannot be realized.
The particle counting and detecting device comprises a detection chip for detecting particles, a signal amplification module for amplifying an electric signal generated after the detection chip detects the particles, and a signal acquisition and analysis module for acquiring the signal amplified by the signal amplification module and counting particles with different sizes respectively;
the detection chip is formed by bonding an ITO conductive glass substrate and a PDMS micro-current control chip;
the ITO glass substrate is provided with an ITO driving electrode group serving as a driving electrode and an ITO detection electrode group serving as a detection electrode;
the ITO driving electrode group is connected with an external power supply through an interface I and an interface II;
the ITO drive electrode group comprises an ITO drive electrode anode and an ITO drive electrode cathode;
the ITO detection electrode group is connected with the signal amplification module through an interface III and an interface IV;
the ITO detection electrode group comprises an ITO detection electrode anode and an ITO detection electrode cathode;
the ITO drive electrode group and the ITO detection electrode group extend to the edge of the ITO glass substrate;
one side of the PDMS microfluidic chip is provided with a concave micro-channel;
the micro-channel comprises a main branch channel for mixing particles and sheath liquid, a particle lateral branch channel for guiding the particles to enter the main branch channel, a sheath liquid lateral branch channel for guiding the sheath liquid to enter the main branch channel, and a detection lateral branch channel for guiding small particles to flow out;
one end of each of the particle lateral branch channel, the sheath fluid lateral branch channel and the detection lateral branch channel is communicated with the main branch channel;
the other ends of the particle side branch channel, the sheath fluid side branch channel, the detection side branch channel and the main branch channel are respectively provided with a particle inlet well, a sheath fluid inlet well, a small particle outlet well and a large particle outlet well;
the tail end of the detection side branch channel is folded inwards in a V shape and is connected with the main branch channel, and a small opening of the detection side branch channel, which is connected with the main branch channel, is a detection opening;
the tail ends of the ITO driving electrode anode and the ITO driving electrode cathode are respectively positioned below the sheath fluid inlet well and the large-particle outlet well;
the ITO detection electrode positive pole and the ITO detection electrode negative pole are located the detection mouth both sides respectively, and ITO detection electrode negative pole end is located under the detection mouth.
Furthermore, the number of the detection side branch channels is any number, namely the detection side branch channels comprise a first detection side branch channel, a second detection side branch channel and an Nth detection side branch channel;
the distance between the first detection side branch channel and the particle side branch channel is at least 2cm, and the rest detection side branch channels are arranged at equal intervals in sequence.
Further, the particle inlet well, the sheath fluid inlet well, the small particle outlet well and the large particle outlet well all have diameters of 2-3 mm.
Further, the width of the detection port is adjusted according to the number of the detection side branch channels.
Further, the depth and width ratios of the particle side branch channel, the sheath fluid side branch channel and the detection side branch channel to the main branch channel are all 1: 2.
Further, the distance between the cathode end of the ITO detection electrode and the detection port is about 1/2 of the width of the main branch channel.
A process for the detection of particles in a wide range of sizes comprising the steps of:
s1: infiltrating a channel; firstly, injecting buffer solution into each channel of a detection chip by using a pipette through a sheath solution inlet well, placing the detection chip on a horizontal desktop, standing for 1-2 minutes, and after the buffer solution infiltrates the whole channel, sequentially and uniformly injecting the buffer solution into all inlet and outlet wells by using the pipette;
s2: adding a reagent; connecting an injector I filled with sheath fluid solution to a sheath fluid inlet well, connecting an injector II filled with to-be-detected particle solution to a particle inlet well, and respectively fixing the injector I and the injector II on two micro injection pumps;
s3: a connection circuit; respectively connecting the positive electrode of the ITO driving electrode and the negative electrode of the ITO driving electrode to the positive electrode and the negative electrode of a direct-current 6V power supply; connecting an ITO detection electrode positive electrode and an ITO detection electrode negative electrode with positive and negative leads of a signal amplification module through a special clamp pin for an ITO conductive glass substrate;
s4: debugging the flow rate; opening the two micro injection pumps, adjusting the flow rates of the two micro injection pumps to 0.5ml/h, pressing a start button of the two micro injection pumps, after the two injection pumps stably operate for 1-2 minutes, firstly adjusting the flow rate of the micro injection pump fixedly provided with the syringe I for containing the sheath solution to be detected to be 1.5-2 ml/h, then adjusting the flow rate of the micro injection pump fixedly provided with the syringe II for containing the particle solution to be detected to be 0.17-0.2 ml/h, and enabling the flow rate ratio of the micro injection pump fixedly provided with the syringe I for containing the sheath solution to be detected to the micro injection pump fixedly provided with the syringe II for containing the particle solution to be detected to be about 8; stably operating the micro injection pump of the injector containing the sheath solution and the micro injection pump of the injector containing the particle solution to be detected for 1-2 minutes;
s5: detecting a signal; opening a signal acquisition card of the signal acquisition and analysis module, and opening a matched Labview signal acquisition program; voltage pulse signals of particles flowing through two ends of the detection port are collected through an ITO detection electrode group, collected information is amplified through a signal amplification module, and is processed by a signal collection and analysis module to be recorded and display corresponding detection data, namely the number of detection sample particles; the signal to noise ratio of the signal is improved by adjusting the flow rates of the two micro injection pumps when the signal is collected; the signal data is recorded and saved.
Further, the concentration of the sheath solution is less than the concentration of the particle solution.
Further, the flow rate ratio of the micro injection pump of the syringe containing the sheath solution to the micro injection pump of the syringe containing the particle solution to be detected is not less than 8.
When the detection particles flow through the detection port, the particles with the particle size smaller than the size of the detection door enter the detection channel, the particles with the particle size larger than the size of the detection door continuously flow forwards after passing through the detection port, and both the particles can cause the change of an electric field near the detection port, so that detection signals can be generated, the amplitudes of the two detection signals are relatively close, real-time observation can be realized, based on an impedance pulse sensing detection technology, the accurate detection of the particles in a wide size range is realized by utilizing the flow inertial lift force and simultaneously adopting two modes of 'passing the particles through the detection port' and 'not passing through the detection port', and the particles in the range of 0.5-50 mu m can be accurately detected by using the device provided by the invention.
Drawings
In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings needed to be used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments described in the present application, and other drawings can be obtained by those skilled in the art without creative efforts.
FIG. 1 is a schematic view of the overall structure of the apparatus of the present invention;
FIG. 2 is a schematic diagram of the structure of an ITO electrode of the device of the present invention;
FIG. 3 is a schematic diagram of the structure of PDMS sheet of the device of the present invention.
In the figure: 1. the device comprises a detection chip, 2, a signal amplification module, 3, a signal acquisition and analysis module, 4, an ITO conductive glass substrate, 5, a PDMS micro-current control chip, 6, an ITO driving electrode anode, 7, an ITO driving electrode cathode, 8, interfaces I, 9, interfaces II, 10, interfaces III, 11, interfaces IV, 12, an ITO detection electrode anode, 13, an ITO detection electrode cathode, 14, a main branch channel, 15, a particle side branch channel, 16, a sheath fluid side branch channel, 17, a detection side branch channel, 18, a particle inlet well, 19, a sheath fluid inlet well, 20, a small particle outlet well, 21, a large particle outlet well, 22 and a detection port.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention clearer, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all embodiments of the present invention. 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.
The sample flow is extruded by using buffer solution (sheath fluid), so that the sample flow is focused at a narrow width, and particles are pushed to different flow lines by the aid of the adjusting device and various parameters of the fluid, such as flow ratio, solution viscosity, channel depth-to-width ratio and the like, through induced inertial lift force, and then enter different detection ports in batches, and particle detection is achieved.
When the particles flow through the micropores with the electric field, the particles can generate obvious disturbance to the electric field, and simultaneously, the voltage at two ends of the corresponding micropores is changed to generate a voltage pulse signal. Based on the principle, particles can flow through the detection micropores, voltage pulse signals generated at two ends of the micropores are detected by using the ITO electrodes and are output to the NI data acquisition card through the amplification circuit, and NI output signals can be directly displayed and analyzed on a connected computer.
Fig. 1 is a schematic diagram of the overall structure of the device of the present invention, fig. 2 is a schematic diagram of the structure of an ITO electrode of the device of the present invention, fig. 3 is a schematic diagram of the structure of a PDMS sheet layer of the device of the present invention, and a particle detection device with a wide size range includes:
the particle counting and detecting device comprises a detection chip 1 for detecting particles, a signal amplification module 2 for amplifying an electric signal generated after the detection chip 1 detects the particles, and a signal acquisition and analysis module 3 for acquiring the signal amplified by the signal amplification module 2 and counting particles with different sizes respectively;
the ITO conductive glass substrate 4 and the PDMS micro-current control chip 5 are irreversibly bonded to form a detection chip 1 based on impedance pulse sensing and with particles in a wide size range after being treated by a plasma cleaner;
before the ITO glass substrate 4 is treated by a plasma cleaning machine, the ITO glass substrate 4 is firstly put into an acetone solution to be soaked for more than 4 hours, then the glass substrate 4 filled with the acetone solution and the ITO glass substrate is easily put into an ultrasonic cleaning machine to be cleaned for more than 5 minutes, after the ITO glass substrate 4 is taken out by tweezers, absolute ethyl alcohol is firstly used for washing the acetone, then pure water is used for washing the absolute ethyl alcohol, then the nitrogen is used for drying the pure water, and the pure water is put into the plasma cleaning machine; the PDMS micro-control flow chip 5 is firstly punched at the positions of the particle inlet well 18, the sheath fluid inlet well 19, the small particle outlet well 20 and the large particle outlet well 21 by a puncher, then the residual PDMS scraps in the particle inlet well 18, the sheath fluid inlet well 19, the small particle outlet well 20 and the large particle outlet well 21 are blown by nitrogen, and then the PDMS micro-control flow chip is placed in a plasma cleaning machine.
The ITO glass substrate 4 is provided with an ITO driving electrode group serving as a driving electrode and an ITO detection electrode group serving as a detection motor;
the ITO driving electrode group is connected with an external power supply through an interface I8 and an interface II 9;
the ITO drive electrode group comprises an ITO drive electrode anode 6 and an ITO drive electrode cathode 7;
the ITO detection electrode group is connected with the signal amplification module 2 through an interface III 10 and an interface IV 11; the ITO detection electrode group comprises an ITO detection electrode anode 12 and an ITO detection electrode cathode 13;
the ITO drive electrode group and the ITO detection electrode group both extend to the edge of the ITO glass substrate 4;
one surface of the PDMS microfluidic chip 5 is provided with a concave micro-channel;
the micro-channel comprises a main branch channel 14 for mixing particles and sheath liquid, a particle side branch channel 15 for guiding large particles to enter the main branch channel 14, a sheath liquid side branch channel 16 for guiding the sheath liquid to enter the main branch channel 14, and a detection side branch channel 17 for guiding small particles to enter a small particle outlet well 20;
the particle lateral branch channel 15, the sheath fluid lateral branch channel 16 and the detection lateral branch channel 17 are all communicated with the main branch channel 14;
the other ends of the particle side branch channel 15, the sheath fluid side branch channel 16, the detection side branch channel 17 and the main branch channel 14 are respectively provided with a particle inlet well 18, a sheath fluid inlet well 19, a small particle outlet well 20 and a large particle outlet well 21; the small particle outlet well 20 and the large particle outlet well 21 are used for storing waste liquid;
the tail end of the detection side branch channel 17 is folded inwards in a V shape and is connected with the main branch channel 14, and a small opening of the detection side branch channel 17, which is connected with the main branch channel 14, is a detection opening 22;
the particle side branch channel 15 and the sheath fluid side branch channel 16 are both 4mm in length and 50 μm in width, the detection side branch channel 17 is 8mm in length and 300 μm in width, the main branch channel 17 is 3cm in length and 50 μm in width, the detection port 22 is 2 μm in width, and all the channels are 25 μm in height; the diameters of the particle inlet well 18, the sheath fluid inlet well 19, the small particle outlet well 20 and the large particle outlet well 21 are all 2mm, the solvent is glycerol aqueous solution, the buffer solution is deionized water, and the electric field intensity applied to the anode and the cathode of the channel is 40V/cm.
According to different practical situations, if a plurality of detection side branch channels exist, the widths of the detection ports corresponding to different detection side branch channels are sequentially increased from any minimum value to any maximum value by a proper value and the like, and the narrower the width of the detection port is, the closer the detection port is to the initial end of the main channel.
The tail ends of the ITO driving electrode anode 6 and the ITO driving electrode cathode 7 are respectively positioned below the sheath fluid inlet well 19 and the large-particle outlet well 21;
the ITO detection electrode anode 12 and the ITO detection electrode cathode 13 are respectively positioned at two sides of the detection port 22, and the tail end of the ITO detection electrode cathode 13 is positioned under the detection port 22.
The method for counting particles based on the particle counting device takes polypropylene sample particles with different particle sizes as an example for detection, and specifically comprises the following steps:
s1: soaking the channel, namely firstly injecting ionized water into a channel of the detection chip 1 through a sheath fluid inlet well 19 by using a pipette, placing the PDMS micro-flow control chip on a horizontal desktop, standing for 1-2 minutes, and after the whole channel is soaked by the deionized water, sequentially and uniformly injecting the ionized water into all inlet and outlet wells by using the pipette;
s2: introducing a solution, connecting an injector I filled with deionized water (sheath fluid) to a sheath fluid inlet well 19 by using a Teflon hard tube, connecting an injector II filled with a particle solution to be detected to a particle inlet well 18 by using a Teflon hard tube, and respectively fixing the injector I and the injector II on two micro injection pumps;
s3: a connection circuit; respectively connecting an ITO driving electrode anode 6 and an ITO driving electrode cathode 7 to the anode and the cathode of a direct current 6V power supply; connecting an ITO detection electrode anode 12 and an ITO detection electrode cathode 13 to a collecting anode and a collecting cathode of the signal amplification module 2, wherein both groups of electrodes are connected with a common lead by using a special ITO conductive glass clamp needle; the signal amplification module 2 is connected with a signal acquisition card, and the signal acquisition card is connected with a computer;
s4: debugging the flow rate; opening the two micro injection pumps, adjusting the flow rates of the two micro injection pumps to 0.5ml/h, pressing a start button of the two micro injection pumps, after the two injection pumps stably run for 1-2 minutes, firstly adjusting the flow rate of the micro injection pump fixedly provided with an injector I for containing deionized water to be 1.5-2 ml/h, then adjusting the flow rate of the micro injection pump fixedly provided with an injector II for containing a particle solution to be detected to be 0.17-0.2 ml/h, and enabling the flow rate ratio of the two micro injection pumps to be 8; after the two micro-injection pumps stably operate for 1-2 minutes;
s5: detecting a signal; opening a signal acquisition card; opening a matched Labview signal acquisition program; voltage pulse signals generated when particles flow through the detection port 22 are collected through the ITO electrode, collected information is amplified through the signal amplification module 2, is processed through the signal collection and processing module 3, is recorded, and displays corresponding detection data, namely the number of detected sample particles, and the signal to noise ratio of the signals is improved by adjusting the flow rate of the two micro-injection pumps when the signals are collected; recording and storing the signal data;
the specific detection result is as follows: real-time detection and counting results can be directly obtained through a computer, the number of particles in a sample is equal to the number of effective pulse signals, and the counting results of all detection ports can be respectively displayed on different counting plates or distinguished through lines with different colors on the same counting panel.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (9)
1. A wide size range's particle detection device which characterized in that: the method comprises the following steps:
the particle counting and detecting device comprises a detection chip (1) for detecting particles, a signal amplification module (2) for amplifying an electric signal generated after the detection chip (1) detects the particles, and a signal acquisition and analysis module (3) for acquiring the signal amplified by the signal amplification module (2) and counting particles with different sizes respectively;
the detection chip (1) is formed by bonding an ITO conductive glass substrate (4) and a PDMS micro-current control chip (5);
the ITO glass substrate (4) is provided with an ITO driving electrode group serving as a driving electrode and an ITO detection electrode group serving as a detection electrode;
the ITO driving electrode group is connected with an external power supply through an interface I (8) and an interface II (9);
the ITO drive electrode group comprises an ITO drive electrode anode (6) and an ITO drive electrode cathode (7);
the ITO detection electrode group is connected with the signal amplification module (2) through an interface III (10) and an interface IV (11);
the ITO detection electrode group comprises an ITO detection electrode anode (12) and an ITO detection electrode cathode (13);
the ITO driving electrode group and the ITO detection electrode group both extend to the edge of the ITO glass substrate (4);
one surface of the PDMS microfluidic chip (5) is provided with a micro-channel which is concavely engraved;
the micro-channel comprises a main branch channel (14) for mixing particles and sheath liquid, a particle side branch channel (15) for guiding the particles to enter the main branch channel (14), a sheath liquid side branch channel (16) for guiding the sheath liquid to enter the main branch channel (14), and a detection side branch channel (17) for guiding small particles to flow out;
one end of each of the particle lateral branch channel (15), the sheath fluid lateral branch channel (16) and the detection lateral branch channel (17) is communicated with the main branch channel (14);
the other ends of the particle side branch channel (15), the sheath fluid side branch channel (16), the detection side branch channel (17) and the main branch channel (14) are respectively provided with a particle inlet well (18), a sheath fluid inlet well (19), a small particle outlet well (20) and a large particle outlet well (21);
the tail end of the detection side branch channel (17) is folded inwards in a V shape and is connected with the main branch channel (14), and a small opening of the detection side branch channel (17) connected with the main branch channel (14) is a detection opening (22);
the tail ends of the ITO driving electrode positive electrode (6) and the ITO driving electrode negative electrode (7) are respectively positioned below the sheath fluid inlet well (19) and the large-particle outlet well (21);
the ITO detection electrode anode (12) and the ITO detection electrode cathode (13) are respectively positioned at two sides of the detection port (22), and the tail end of the ITO detection electrode cathode (13) is positioned under the detection port (22).
2. A wide range of particle detection apparatus as claimed in claim 1, wherein: the number of the detection side branch channels (17) is any, namely the detection side branch channels (17) comprise a first detection side branch channel, a second detection side branch channel and an Nth detection side branch channel;
the distance between the first detection side branch channel and the particle side branch channel (15) is at least 2cm, and the rest detection side branch channels are arranged at equal intervals in sequence.
3. A wide range of particle detection apparatus as claimed in claim 1, wherein: the diameters of the particle inlet well (18), the sheath fluid inlet well (19), the small particle outlet well (20) and the large particle outlet well (21) are all 2-3 mm.
4. A wide range of particle detection apparatus as claimed in claim 1, wherein: the width of the detection port (22) is adjusted according to the number of the detection side branch channels (17).
5. A wide range of particle detection apparatus as claimed in claim 1, wherein: the depth and width ratios of the particle side branch channel (15), the sheath fluid side branch channel (16) and the detection side branch channel (17) to the main branch channel (14) are all 1: 2.
6. A wide range of particle detection apparatus as claimed in claim 1, wherein: the distance between the tail end of the ITO detection electrode cathode (13) and the detection port (22) is about 1/2 of the width of the main branch channel (14).
7. A process for the detection of particles of a broad size range as claimed in any one of claims 1 to 6, further characterized by: the method comprises the following steps:
s1: infiltrating a channel; firstly, injecting buffer solution into each channel of a detection chip (1) by using a pipette through a sheath solution inlet well (19), placing the detection chip on a horizontal desktop, standing for 1-2 minutes, and after the buffer solution infiltrates the whole channel, sequentially and uniformly injecting the buffer solution into all inlet and outlet wells by using the pipette;
s2: adding a reagent; connecting an injector I filled with sheath fluid solution to a sheath fluid inlet well (19), connecting an injector II filled with to-be-detected particle solution to a particle inlet well (18), and respectively fixing the injector I and the injector II on two micro injection pumps;
s3: a connection circuit; respectively connecting an ITO driving electrode positive electrode (6) and an ITO driving electrode negative electrode (7) to a positive electrode and a negative electrode of a direct-current 6V power supply; an ITO detection electrode positive electrode (12) and an ITO detection electrode negative electrode (13) are connected with positive and negative leads of a signal amplification module (2) through a special clamp needle of an ITO conductive glass substrate (4);
s4: debugging the flow rate; opening the two micro injection pumps, adjusting the flow rates of the two micro injection pumps to 0.5ml/h, pressing a start button of the two micro injection pumps, after the two injection pumps stably operate for 1-2 minutes, firstly adjusting the flow rate of the micro injection pump fixedly provided with the syringe I for containing the sheath solution to be detected to be 1.5-2 ml/h, then adjusting the flow rate of the micro injection pump fixedly provided with the syringe II for containing the particle solution to be detected to be 0.17-0.2 ml/h, and enabling the flow rate ratio of the micro injection pump fixedly provided with the syringe I for containing the sheath solution to be detected to the micro injection pump fixedly provided with the syringe II for containing the particle solution to be detected to be about 8; stably operating the micro injection pump of the injector containing the sheath solution and the micro injection pump of the injector containing the particle solution to be detected for 1-2 minutes;
s5: detecting a signal; opening a signal acquisition card of the signal acquisition and analysis module (3), and opening a matched Labview signal acquisition program; voltage pulse signals of particles flowing through two ends of a detection port (22) are collected through an ITO detection electrode group, collected information is amplified through a signal amplification module (2), and is processed by a signal collection and analysis module (3) to be recorded and display corresponding detection data, namely the number of detected sample particles; the signal to noise ratio of the signal is improved by adjusting the flow rates of the two micro injection pumps when the signal is collected; the signal data is recorded and saved.
8. A process for the detection of particles of a broad size range as defined in claim 7, wherein: the concentration of the sheath fluid solution is less than the concentration of the particle solution.
9. A process for the detection of particles of a broad size range as defined in claim 7, wherein: the flow rate ratio of the micro injection pump of the injector for containing the sheath solution to the micro injection pump of the injector for containing the particle solution to be detected is more than or equal to 8.
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