CN106644900A - Pulse impedance particle counting device based on non-uniform electric field and particle counting method - Google Patents
Pulse impedance particle counting device based on non-uniform electric field and particle counting method Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 47
- 239000002245 particle Substances 0.000 title claims abstract description 47
- 230000005684 electric field Effects 0.000 title abstract description 12
- 238000001514 detection method Methods 0.000 claims abstract description 48
- 239000004205 dimethyl polysiloxane Substances 0.000 claims abstract description 10
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 claims abstract description 10
- 239000011521 glass Substances 0.000 claims abstract description 9
- 239000007788 liquid Substances 0.000 claims description 64
- 238000003860 storage Methods 0.000 claims description 62
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 claims description 28
- 230000003321 amplification Effects 0.000 claims description 15
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 15
- 229910052697 platinum Inorganic materials 0.000 claims description 14
- 238000012360 testing method Methods 0.000 claims description 14
- SNRUBQQJIBEYMU-UHFFFAOYSA-N dodecane Chemical compound CCCCCCCCCCCC SNRUBQQJIBEYMU-UHFFFAOYSA-N 0.000 claims description 12
- 230000008569 process Effects 0.000 claims description 12
- 235000013870 dimethyl polysiloxane Nutrition 0.000 claims description 7
- 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 description 7
- 238000004987 plasma desorption mass spectroscopy Methods 0.000 claims description 7
- 238000004458 analytical method Methods 0.000 claims description 6
- 239000012530 fluid Substances 0.000 claims description 5
- 230000000694 effects Effects 0.000 claims description 3
- 230000003139 buffering effect Effects 0.000 claims description 2
- 230000005611 electricity Effects 0.000 claims description 2
- 238000002347 injection Methods 0.000 abstract 4
- 239000007924 injection Substances 0.000 abstract 4
- -1 polydimethylsiloxane Polymers 0.000 abstract 1
- 230000008859 change Effects 0.000 description 11
- 239000000243 solution Substances 0.000 description 10
- 238000005516 engineering process Methods 0.000 description 3
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- 239000004793 Polystyrene Substances 0.000 description 2
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- 238000011161 development Methods 0.000 description 2
- 235000008434 ginseng Nutrition 0.000 description 2
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- 238000012544 monitoring process Methods 0.000 description 2
- 239000002105 nanoparticle Substances 0.000 description 2
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- 229920002223 polystyrene Polymers 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
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- IXSZQYVWNJNRAL-UHFFFAOYSA-N etoxazole Chemical compound CCOC1=CC(C(C)(C)C)=CC=C1C1N=C(C=2C(=CC=CC=2F)F)OC1 IXSZQYVWNJNRAL-UHFFFAOYSA-N 0.000 description 1
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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
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/1023—Microstructural devices for non-optical measurement
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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
- G01N2015/1024—Counting particles by non-optical means
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Abstract
The invention discloses a pulse impedance particle counting device based on non-uniform electric field and a particle counting method; the pulse impedance particle counting device comprises a glass baseplate, a PDMS (polydimethylsiloxane) microfluidic chip, a signal amplifying element and a signal collection control system; the PDMS microfluidic chip is provided with a microchannel by means of indenting, and the microchannel includes: a main channel with two ends provided with an oil incoming liquid-storing hole and an oil outgoing liquid-storing hole respectively; a sample injection channel extending from the middle of the main channel to a direction leaving the main channel, wherein the tail end of the sample injection channel is provided with a sample injection channel liquid-storing hole; a detection channel extending from a position, a certain distance away from an intersection of the main channel and the sample injection channel, to a direction leaving the main channel, wherein a ratio of the width of the detection channel to the width of the main channel is fixed, and the tail end of the detection channel is provided with a detection channel liquid-storing hole. The pulse impedance particle counting device is simple in structure and high in detection precision, and can provide detection and counting with no need for particles to accessing a detection area.
Description
Technical field
The present invention relates to particle monitoring technique field, particularly relates to a kind of impedance pulse based on inhomogeneous field
Grain counting device and its method of counting.
Background technology
Particle in sample is counted exactly is had great importance and demand in multiple fields；For example, in biology
Medical research, public health detection and the field such as marine environmental monitoring, to research for determine object (as bacterium, virus and
Marine microorganism etc.) number portable particle fast counting apparatus or method, have urgent demand always.
The method of existing conventional grain count at present includes following several：
1) light blockage method (principle of optical interference)：Light blockage method is the grain count method being most widely used at present, also known as light
Obstruction method or light block method, are to carry out the side of Particle Sizing to the occurred intensity variation of blocking of light using particulate
Method.The method principle is simple, but needs costly and baroque photo-translating system, and accuracy of detection is low；
2) inductance method：Inductance method can change the phenomenon of micropore inductance when being passed through and detected micropore using particle, to detecting micropore
The change of inductance signal is monitored, it is possible to achieve grain count.The method principle is simple, and various metallic particles can be carried out
Detection, but metallic particles is only applicable to, accuracy of detection is also limited；
3) capacitance method：Capacitance method is a kind of non-contact detection method, when particle is through detection micropore, can change micropore
Electric capacity, by monitor micropore capacitance signal change, it is possible to achieve grain count.The method can be used for low conducting solution
Grain count in (such as fluid), and capacitance detecting object is mainly metallic particles, but the detection of small capacitance depends on height
The detecting instrument of precision, thus limit its portability development.
4) laser-Induced Fluorescence Detection method：Particle carries fluorescence after laser irradiation, can be detected by photoelectric sensor
And realize grain count.The method is particularly suited for blood analysis, immunology and microbiology association area, counts accurate, inspection
Survey convenient, but sample must be processed before detection, optical element high cost and complex structure used by detection, it is impossible to detect
Atomic little particle；
5) resistance impulse method (RPS)：Resistance impulse method is based on when particle flows through the micropore that there is electric field, two ends at micropore
Voltage change and produce detection signal, realize detection；The method is easy to operate, is current precision highest grain count method.
Micro flow control chip device can be by biology, chemistry, the sample preparation of medical analysis process, reaction, separation and detection
It is integrated on the chip of one piece of micro-meter scale Deng basic operation, is automatically performed analysis overall process.It has liquid flowing it is controllable, disappear
The features such as consumption reagent and sample are few, analyze speed tenfold hundreds of times ground is improved, can be in a few minutes or even shorter time
Inside carry out being analyzed while up to a hundred samples, and pretreatment and the analysis overall process of energy canbe used on line sample.
With the fast development of micro-fluidic chip process technology in recent years, at present existing many is utilized on micro-fluidic chip
The research report that RPS methods are counted to micro-nano particle, cell, bacterium and virus etc..Such as, typical micro-fluidic RPS counts core
Piece is to process one section of live width sense channel more slightly larger than particle to be detected in the middle part of the wide main channel of hundreds of micron, then
Apply a DC electric field at main channel two ends；When the insulated particle in solution is through sense channel its two ends component voltage can become
Change, by obtaining a pulse signal using suitable signal acquiring system, the number of pulse signal is the number of particle.For
The convenient change for obtaining the polygonal voltage of sense channel two, people have invented different chip structures.
Initial detection chip structure only includes the wide main channel of hundreds of micron and be arranged in the middle part of main channel one
The section live width sense channel more slightly larger than candidate particles, by will be inserted in after the platinum electrode series resistance at main channel two ends and direct current
Source two ends are connected, and realize grain count by amplifying the change of resistance both end voltage signal, but the method system noise is big,
Signal to noise ratio is low, so accuracy of detection is low；Detection chip later respectively set up a detection arm passage before and after sense channel, use
Electrode direct measurement passage both end voltage signal, then by differential amplification, it is improved detection signal-to-noise ratio, and then cause detection
Precision is raised；Newest counting chip employs the structure of public inlet opening multichannel annular spread, and every passage all includes master
Passage, sense channel and liquid storage hole etc., its inlet opening connection positive source, after each liquid storage hole series resistance power cathode is connected,
Mutually as reference, current counting chip can preferably eliminate system noise to each passage, by controlling each passage work successively
Make, realize that high flux is counted.It should be understood that, current counting chip is past in order to obtain higher detection signal-to-noise ratio
Toward it is required that the adjoining dimensions of sense channel size and particle, then corresponding to the detection of nano particle also needs accordingly to process
The sense channel of nano-scale, but this is accomplished by the process equipment of extremely complex procedure of processing and costliness.
Further, since the size and particle of sense channel are closer to, particle occlusion detection passage is also easily caused；Together
When, the part that the impurity in solution will also result in sense channel is even completely plugged so that missing occurs in grain diameter testing result
Difference detects interruption, have impact on precision and the stable operation of system.
The content of the invention
In view of the deficiency that prior art is present, the invention aims to provide a kind of impedance arteries and veins based on inhomogeneous field
Grain count device is rushed, in detection process, the candidate particles enter main channel from sample channel, then flow through main channel and sample
Product passage intersection, i.e. particle do not enter sense channel, so as to effectively increase accuracy of detection and anti-channel block ability.
To achieve these goals, technical solution of the present invention is as follows：
A kind of impedance pulse grain count device based on inhomogeneous field, it includes the micro-fluidic core of glass negative, PDMS
Piece, signal amplification component and signal acquisition control system, the PDMS micro-fluidic chips intaglio has the side of microchannel and institute
State glass negative to be packaged as a whole, to form the microchannel for testing sample circulation, it is characterised in that the microchannel includes：
It is respectively arranged at two ends with the main channel of oil-feed liquid storage hole and fuel-displaced liquid storage hole；
From the main channel centre position, to the sample intake passage that the direction away from the main channel extends, the sample intake passage
End is provided with sample intake passage liquid storage hole；
From apart from the position of the main channel and sample intake passage intersection certain distance, to away from the main channel
The sense channel that direction extends, the width of the sense channel is fixed with the width ratio of main channel and end is provided with sense channel
Liquid storage hole；
Simultaneously platinum electrode is inserted in the sample intake passage liquid storage hole and sense channel liquid storage hole, in the sample intake passage hole
Platinum electrode be connected with the positive pole of dc source by a reference resistance, the platinum electrode in the sense channel liquid storage hole with it is above-mentioned
The negative pole connection of dc source；The two ends of the reference resistance are connected by wire with the input of the signal amplification component
Connect, the output end of the signal amplification component is connected with the signal acquisition control system.
Further, as the preferred of the present invention：
The sense channel is angled with main channel, and it preferably adopts 1/200 with the width ratio of main channel.
Further, as the preferred of the present invention：
The signal amplification component adopts difference amplifier element.
Further, as the preferred of the present invention：
The signal acquisition control system includes NI capture cards and computer.
Another object of the present invention is to provide for a kind of method for carrying out grain count based on above-mentioned grain count device, and it is special
Levy and be, comprise the steps：
1) sample is added dropwise：First a certain amount of PBS is added dropwise in the sample intake passage liquid storage hole and sense channel liquid storage hole
Buffer solution, in main channel oil-feed liquid storage hole a certain amount of dodecane is added dropwise, and then a certain amount of testing sample is added dropwise to into institute
In stating sample intake passage liquid storage hole；
2) sample transport：Connect the dc source so that the testing sample in the sample intake passage liquid storage hole is in electric osmose
The main channel is transported under stream and pressure effect, the fuel-displaced liquid storage hole of main channel is then flowed under the pressure differential of main channel；
3) signal amplifies collection analysis：By the platinum electrode collection ginseng in sense channel liquid storage hole and sample intake passage liquid storage hole
Examine the voltage pulse signal at resistance two ends, the signal for collecting after the difference amplifier that is connected with reference resistance amplifies, by
Signal acquisition control system is recorded and is shown corresponding detection data, the i.e. number of detection sample particle；Aforementioned described
The number detected value of grain is equal to the number of sense channel pulse signal.
Particle solution is focused on and flows through main channel and sense channel intersection by dodecane used in detection process.
Compared with prior art, beneficial effects of the present invention：
1) when being detected, its testing sample particle does not enter sense channel to the present invention, blocks up so as to not result in passage
The phenomenon of plug, detection stability is high；
2) present invention improves constantly the signal to noise ratio of detection signal by increasing main channel with the ratio of sense channel, so as to
Improve accuracy of detection.
Description of the drawings
Fig. 1 is the microfluidic chip structure schematic diagram of counting device of the present invention；
Fig. 2 is present system structure chart.
In figure：M, micro-fluidic chip, L, glass negative, A, main channel oil-feed liquid storage hole, B, the fuel-displaced liquid storage hole in main channel, C,
Sample intake passage liquid storage hole, D, sense channel liquid storage hole, 1, main channel, 2, sample intake passage, 3, sense channel.
Specific embodiment
To make the object, technical solutions and advantages of the present invention clearer, below in conjunction with the embodiment of the present invention in it is attached
Figure, is clearly and completely described to technical scheme, it is clear that described embodiment is that a part of the invention is real
Apply example, rather than the embodiment of whole.Based on the embodiment in the present invention, those of ordinary skill in the art are not making creation
Property work under the premise of the every other embodiment that obtained, belong to the scope of protection of the invention.
When particle flows through the micropore that there is electric field, significantly disturbance, both end voltage at corresponding micropore can be produced to electric field
Change and produce voltage pulse signal.The number of detection voltage pulse signal, you can realize the number of detection sample particle.
Based on above-mentioned design background, the present invention devises a kind of impedance pulse grain count device based on inhomogeneous field
And its method of counting, below in conjunction with the accompanying drawings and specific embodiment further illustrates technical scheme：
As shown in figure 1, a kind of impedance pulse grain count device based on inhomogeneous field, it include glass negative L,
PDMS micro-fluidic chip M, signal amplification component and signal acquisition control system.
The PDMS micro-fluidic chips intaglio has the side of microchannel and the glass negative to be packaged as a whole, to form confession
The microchannel of testing sample circulation, it is characterised in that the microchannel includes：It is respectively arranged at two ends with oil-feed liquid storage hole and fuel-displaced storage
The main channel of fluid apertures；From the main channel centre position, to the sample intake passage that the direction away from the main channel extends, the sample introduction
Channel end is provided with sample intake passage liquid storage hole, and the width of the sample intake passage is fixed with the width ratio of main channel, preferably
1/10；From apart from the position of the main channel and sample intake passage intersection certain distance, to the main channel into certain
The sense channel that the direction of angle extends, the preferably width of the sense channel are 1/200 with the width ratio of main channel, and end
End is provided with sense channel liquid storage hole.
Platinum electrode is inserted in the sample intake passage liquid storage hole and sense channel liquid storage hole.In the sample intake passage liquid storage hole
Platinum electrode be connected with the positive pole of dc source by a reference resistance, the platinum electrode in the sense channel liquid storage hole with it is above-mentioned
The negative pole connection of dc source, to realize that testing sample flows into the electric osmose of main channel from sample intake passage liquid storage hole from sample intake passage
Transportation；Sample is flowed into behind main channel, and the fuel-displaced liquid storage hole in main channel is flowed under pressure differential；
The two ends of the reference resistance are connected by wire with the input of the signal amplification component.Preferably, institute
Signal amplification component is stated using difference amplifier element.
The output end of the signal amplification component is connected with the signal acquisition control system, to realize in sense channel
The amplification acquisition testing process of testing sample particle number signal.Preferably, the signal acquisition control system is gathered including NI
Card and computer.
Detected by taking polystyrene sample particle as an example below, as shown in Figure 2：
The device parameter of the implementation case：Size 1*5 μm (wide × high) of the chip detection passage used by the implementation case,
Distance in the middle of each liquid storage hole to chip main channel is 5cm, main microchannel size be 200*5 μm (wide × high), sample intake passage chi
Very little is 20*5 μm (wide × high)；Detected sample is 3 μm of granules of polystyrene solution；Buffer solution is PBS (1 ×) solution；Apply
In the voltage of sample intake passage liquid storage hole and sense channel liquid storage hole be 48V；
The device includes main channel oil-feed liquid storage hole A, the fuel-displaced liquid storage in main channel being located on the PDMS micro-fluidic chips
Hole B, sample intake passage liquid storage hole C and sense channel liquid storage hole D and main channel 1, sample intake passage 2 and sense channel 3.It is logical in sample introduction
Platinum electrode is inserted in road liquid storage hole C and sense channel liquid storage hole D, and dc source two ends are connected to by resistance R；In resistance R
Two ends are by two conductor in parallel in two inputs of a difference amplifier；The output end of difference amplifier is connected to NI data
The input of capture card；NI output signals directly in the Computer display for being connected and can be analyzed.
Based on the method that above-mentioned grain count device carries out grain count, comprise the steps：
1) sample is added dropwise：A certain amount of PBS bufferings are added dropwise in sample intake passage liquid storage hole and sense channel liquid storage hole first
Liquid, is added dropwise a certain amount of dodecane in the fuel feed hole of main channel, it is ensured that oil liquid pressure is more than always solution pressure, then will be a certain amount of
Testing sample be added dropwise in sample channel inlet opening；
2) sample transport：Dc source is connected, the sample in sample intake passage liquid storage hole C is due to EOF and pressure effect quilt
In being transported to main channel, the fuel-displaced liquid storage hole B in main channel, candidate particles in detection process are then flowed under the pressure differential of main channel
Main channel is entered from sample channel, main channel is then flowed through and is not entered sense channel with sample channel intersection, i.e. particle.Due to
Fluid is non-conductive, and fluid region is not in electric field in passage, therefore, can be limited by controlling the position of oil-water interfaces
The distribution of electric field.In practice, need to ensure distance of the main channel in the oil-water interfaces with sense channel intersection to sense channel
Only 2-3 times of candidate particles particle diameter.
Due to only applying electric field, and the narrower in width of sense channel in sample intake passage and sense channel, therefore detect logical
Road is very big (i.e. power line is very intensive) with the electric-field intensity of main channel juncture area.When particle is handed over through sense channel and main channel
During battery limit (BL) domain, disturbance electric field line can be easy to, make sense channel component voltage change, further result in the electricity at resistance R two ends
Buckling, produces detection signal, and detection signal is input into NI capture cards after the amplification of AD620 difference amplifiers differential signal,
NI capture cards output signal directly can show and be analyzed on the computer for being connected.
3) signal amplifies collection analysis：By the platinum electrode collection ginseng in sense channel liquid storage hole and sample intake passage liquid storage hole
Examine the voltage pulse signal at resistance two ends, the signal for collecting after the difference amplifier that is connected with reference resistance amplifies, by
Signal acquiring system is recorded and is shown corresponding detection data, the i.e. number of detection sample particle.
Testing result：Real-time detection result can be directly obtained by computer, the number of particle is equal to pulse in sample
The number of signal.
Using a kind of impedance pulse grain count device and its method of counting based on inhomogeneous field of the present invention, inspection
During survey, particle solution is focused on using dodecane flows through main channel and sense channel intersection, and the candidate particles from
Sample channel enters main channel, then flows through main channel and sample channel intersection.Due to sense channel and main channel junctional area
The electric-field intensity in domain is very big, and particle does not enter sense channel, so as to effectively increase accuracy of detection and anti-channel block ability.
The above, the only present invention preferably specific embodiment, but protection scope of the present invention is not limited thereto,
Any those familiar with the art the invention discloses technical scope in, technology according to the present invention scheme and its
Inventive concept equivalent or change in addition, all should be included within the scope of the present invention.
Claims (4)
1. a kind of impedance pulse grain count device based on inhomogeneous field, it include glass negative, PDMS micro-fluidic chips,
Signal amplification component and signal acquisition control system, the PDMS micro-fluidic chips intaglio has the side of microchannel and the glass
Glass egative film is packaged as a whole, to form the microchannel for testing sample circulation, it is characterised in that the microchannel includes：
It is respectively arranged at two ends with the main channel of oil-feed liquid storage hole and fuel-displaced liquid storage hole；
From the main channel centre position, to the sample intake passage that the direction away from the main channel extends, the width of the sample intake passage
Degree is fixed with the width ratio of main channel, and end is provided with sample intake passage liquid storage hole；
From apart from the position of the main channel and sample intake passage intersection certain distance, to the direction away from the main channel
The sense channel of extension, the width of the sense channel is fixed with the width ratio of main channel, and end is provided with sense channel storage
Fluid apertures；
Meanwhile, it is inserted with platinum electrode, the sample intake passage liquid storage hole in the sample intake passage liquid storage hole and sense channel liquid storage hole
Interior platinum electrode is connected by a reference resistance with the positive pole of dc source, the platinum electrode in the sense channel liquid storage hole with it is upper
State the negative pole connection of dc source；The two ends of the reference resistance are connected by wire with the input of the signal amplification component
Connect, the output end of the signal amplification component is connected with the signal acquisition control system.
2. grain count device according to claim 1, it is characterised in that：
The sense channel is angled with main channel, and it preferably adopts 1/200 with the width ratio of main channel.
3. a kind of method that grain count device according to claim 1 carries out grain count, it is characterised in that include as
Lower step：
1) sample is added dropwise：A certain amount of PBS bufferings are added dropwise in the sample intake passage liquid storage hole and sense channel liquid storage hole first
Liquid, in main channel oil-feed liquid storage hole a certain amount of dodecane is added dropwise, then by a certain amount of testing sample be added drop-wise to it is described enter
In sample passage liquid storage hole；
2) sample transport：Connect the dc source so that the testing sample in the sample intake passage liquid storage hole in EOF and
The main channel is transported under pressure effect, the fuel-displaced liquid storage hole of main channel is then flowed under the pressure differential of main channel, detected
During candidate particles enter main channel from sample channel, then flow through main channel and sample channel intersection, i.e. particle and do not enter
Enter sense channel；
3) signal amplifies collection analysis：By the platinum electrode collection in sense channel liquid storage hole and sample intake passage liquid storage hole with reference to electricity
The voltage pulse signal at resistance two ends, the signal for collecting after the signal amplification component that is connected with reference resistance amplifies, by believing
Number acquisition control system is recorded and is shown corresponding detection data, the i.e. number of detection sample particle；The aforementioned particle
Number detected value be equal to sense channel pulse signal number.
4. grain count method according to claim 3, it is characterised in that：
Particle solution is focused on and flows through main channel and sense channel intersection by dodecane used in detection process.
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| CN201710108467.1A CN106644900B (en) | 2017-02-27 | 2017-02-27 | Impedance pulse particle counting device based on non-uniform electric field and counting method thereof |
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Cited By (9)
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| CN108226013A (en) * | 2018-04-13 | 2018-06-29 | 大连海事大学 | It is a kind of to improve particle detections precision and the device and method of flux using static focusing |
| CN108279202A (en) * | 2018-03-28 | 2018-07-13 | 大连海事大学 | A kind of adjustable grain count device and method of accuracy of detection |
| CN108458963A (en) * | 2018-04-13 | 2018-08-28 | 大连海事大学 | A kind of micro flow control chip device and method particle being carried out based on nano-micrometre combination of channels and cell sequence is detached and counted |
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