CN105749993A - Microfluidic chip device and microfluidic chip method capable of improving detection precision of resistance-pulse-method particles - Google Patents
Microfluidic chip device and microfluidic chip method capable of improving detection precision of resistance-pulse-method particles Download PDFInfo
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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
- B01L3/502707—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 the manufacture of the container or its components
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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
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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
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
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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
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
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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
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0809—Geometry, shape and general structure rectangular shaped
Abstract
The invention discloses a microfluidic chip device and a microfluidic chip method capable of improving the detection precision of resistance-pulse-method particles. The device is provided with a micro-channel, a particle detection region, a first focusing channel and a second focusing channel, wherein the micro-channel comprises a main channel; the main channel is provided with a first main channel liquid storage hole and a second main channel liquid storage hole; the particle detection region is provided with two detection channels, one end of each detection channel is communicated with the main channel and the other ends of the detection channels are connected with two input ends of a differential amplification element respectively; the output end of the differential amplification element is connected with a display element through a data acquisition unit; a first focusing liquid storage hole is formed in one end of the first focusing channel; a second focusing liquid storage hole is formed in one end of the second focusing channel; and the first main channel liquid storage hole, the second main channel liquid storage hole, the first focusing liquid storage hole and the second focusing liquid storage hole are connected with a direct-current voltage splitting device respectively. According to the microfluidic chip device and microfluidic chip method disclosed by the invention, an effective detection width of a detection region is reduced by adopting focusing sheath liquid with high resistivity, so that the detection precision is improved.
Description
Technical field
The present invention relates to granule detection field, especially relate to a kind of micro flow control chip device that can be effectively improved resistance impulse method granule accuracy of detection and method.
Background technology
Particle monitoring technique has a wide range of applications in such as environmental monitoring, Marine microorganism detection, biological micromolecule detection and clinical diagnosis etc..At present, researcher commonly uses microscope and observes and the means such as image processing techniques are to realize the detection of granule, namely utilizes microscope or imageing sensor to carry out the collection of particle image, then picture signal converts to digital signal input computer processes.This kind of detection means is capable of detecting when particle signal, but its device is mostly expensive, it is long to process the time, and accuracy of detection is limited, is more widely applied thus limiting it.Simultaneously because its detection efficiency of microfluidic chip technology is high, reagent and sample consumption is few and the advantage such as little, being easily integrated of system bulk, thus it is developed rapidly in these fields.
At present, the particle detection technique based on micro-fluidic chip mainly has: shading detection method, fluorescence detection, capacitance detecting method, inductance detection method and resistance detection method etc..Shading detection method utilizes light to block the principle granule to detect in fluid channel；But this detection method is affected relatively big by bubble, impurity etc., and accuracy of detection is not high；Fluorescence detection is the fluorescent grain utilizing laser to irradiate in fluid channel so that it is inspire exciting light, realizes the detection of granule by detecting the intensity of exciting light, but this detection method detection particle type is limited, and equipment is complicated；Capacitance detecting method is the difference utilizing granule with Dielectric constant, the capacitance variation that detection granule causes when flowing through electric capacity, thus realizing the detection of granule, but this detection method detection particle type is limited, capacity arrangement difficulty；When inductance detection method is to utilize ferromagnetic particle to flow through inductance coil, granule reacts on inductance coil after being magnetized so that it is inductance value changes, thus realizing the detection to granule；But this detection method accuracy of detection is not high, and it is only capable of detection ferromagnetic particle；Resistance detection method is application Coulter principle, is flowed through the resistance variations that detection zone causes by granule and the detection realizing granule, the method accuracy of detection is high, easy and simple to handle, and measuring circuit is fairly simple, being easily integrated, therefore it obtains a wide range of applications at micro-fluidic detection field.
Seeing in theory, improving the precision of resistance detection method, namely improve signal to noise ratio, the method that both can pass through to reduce noise realizes, it is also possible to realize by reducing the size of detection zone；Researcher reduces noise already by means such as isolation detection device, signal differential inputs, but to continue reducing noise on this basis, to improve accuracy of detection very difficult.Researcher is by methods such as the nano-pores in use femtosecond laser (femtosecond-pulsedlaser), electron beam lithography, CNT and natural planes lipid bilayer (lipidbilayers), process nano level sense channel, improve accuracy of detection；But above-mentioned nanochannel processing method is expensive, program is complicated, reliability is not high, it is difficult to obtain application in practice.
At present, material and the process of processing micro-fluidic chip are more, and wherein most widely used material is PDMS, and the process of employing is soft lithography.When adopting this technique, the restriction of printing precision during due to printing mask so that the minimum channel size processing chip is limited, namely adopts this processing technique to be only capable of processing the passage being sized larger than 20 microns.When detecting little microparticle or nano-particle when needs, when adopting this micro-fluidic chip, owing to technique restriction can not continue to zoom out detection zone size, detection signal too overly soft pulse will be made to can't detect signal.
Summary of the invention
In view of the defect that prior art exists, the invention aims to provide a kind of micro flow control chip device improving resistance impulse method granule accuracy of detection, this micro flow control chip device is different from sample solution electrical conductivity based on ultra-pure water, ultra-pure water is made to extrude sample solution at micro-fluidic chip granule detection zone by EOF, the effective detection width making detection zone reduces, and then reaches to improve the purpose of accuracy of detection.
To achieve these goals, the technical scheme is that
A kind of micro flow control chip device improving resistance impulse method granule accuracy of detection, this device includes intaglio the PDMS micro-fluidic chip of some fluid channel, described PDMS micro-fluidic chip intaglio has fluid channel side and glass negative sealing-in to be structure as a whole, so that forming microchannel between described PDMS micro-fluidic chip and glass negative, it is characterized in that: described microchannel has for the focus channel flowed in and out focusing on sheath fluid, and microchannel includes
For storing the main channel of sample liquid, this main channel has the first main channel liquid storage hole for the sample liquid before storing detection and for storing the second main channel liquid storage hole of the sample waste after detection；
It is arranged at main channel middle position, the granule detection zone of the detection signal for obtaining and flow through in the analyte sample fluid of one's respective area corresponding to granule, this granule detection zone has two one end and is all connected with described main channel, the detection arm passage that the other end is connected with differential amplification element two input respectively, and this differential amplification cell output is connected with display element by data acquisition element；
And focus channel, this focus channel includes two respectively centered by described granule detection zone, it is symmetricly set in the described granule detection zone left and right sides, for realizing the first focus channel and second focus channel of focussing force, and the first focusing liquid storage hole of detection prefocusing sheath fluid it is arranged to store in first focus channel one end, after second focus channel one end is arranged to storage detection, the second of focusing sheath fluid waste liquid focuses on liquid storage hole；
Described first main channel liquid storage hole, the second main channel liquid storage hole, the first focusing liquid storage hole and second focus on liquid storage hole and are connected with DC partial voltage device respectively, this DC partial voltage device for for described first main channel liquid storage hole, the second main channel liquid storage hole, first focus on liquid storage hole and second and focus on liquid storage hole the voltage of different amplitude is provided, to realize in electric osmose manipulation microchannel testing sample and to drive the flowing focusing on sheath fluid.
Further, described first main channel liquid storage hole, the second main channel liquid storage hole, the first focusing liquid storage hole and second focus on liquid storage hole and are connected with DC partial voltage device respectively through platinum electrode, to overcome employing ordinary electrode, it is easily generated the defect of cotton-shaped pollutant when it is by electrolysis.
Further, each described detection arm passage is connected with differential amplification element two input respectively through platinum electrode.
Further, described data acquisition element adopts Labview data collecting instrument；Described display element adopts PC.
Further, described focusing sheath fluid adopts ultra-pure water.
The present invention also provides a kind of method improving resistance impulse method granule accuracy of detection, and it is realized by aforementioned micro flow control chip device, comprises the steps:
1) in described first main channel liquid storage hole, the second main channel liquid storage hole, add a certain amount of phosphate-PBS buffer solution, focus on first in liquid storage hole and the second focusing liquid storage hole and add a certain amount of ultra-pure water；In the liquid storage hole of detection arm passage, insert platinum electrode respectively, and aforementioned platinum electrode is connected with differential amplification element respectively；
(2) regulating DC partial voltage device makes each output current potential be setting value, and the first corresponding with micro-fluidic chip for each outfan main channel liquid storage hole, the second main channel liquid storage hole, the first focusing liquid storage hole and second are focused on liquid storage hole and be connected；
(3) add a certain amount of detected sample liquid at the first main channel liquid storage hole, add same amount of phosphate buffered solution at the second main channel liquid storage hole, focus on first in liquid storage hole and the second focusing liquid storage hole and add same amount of ultra-pure water；
(4) utilize Labview data collecting instrument and PC, carry out signals collecting, display and storage.
Compared with prior art, beneficial effects of the present invention:
1. the present invention by focus channel inner focusing sheath fluid-ultra-pure water to the squeezing action at granule detection zone of the sample solution in main channel so that the actual size of sample solution greatly reduces, thus smaller granule being detected delicately；And have simple to operate, and in use can adjust the advantages such as focal width neatly；
2. the present invention utilizes DC partial voltage device can be conveniently adjusted output potential value, thus changing the flow regime of main channel and focus channel, and then is conveniently adjusted the sample solution size at detection zone, it is achieved the optimum detection to various little micro-sized particle；
3. the present invention adopts difference amplifier to detect, and weakens signal noise, improves the signal to noise ratio of signal.
Accompanying drawing explanation
Fig. 1 is microfluidic chip structure schematic diagram of the present invention；
Fig. 2 is present configuration schematic diagram；
Fig. 3 is the present invention--embodiment Detection results figure.
In figure: M is PDMS micro-fluidic chip, L is glass negative, and 1 is focus channel, 11, the first focus channel, the 12, first focus channel liquid storage hole, the 13, second focus channel, 14, the second focus channel liquid storage hole, 2 is main channel, the 21, first main channel liquid storage hole, 22, the second main channel liquid storage hole, 3 is detection arm passage, 31, bulge-structure, 4 is DC partial voltage device, and 5 is described micro-fluidic chip, and 6 is differential amplification element, 7 is data acquisition element, and 8 is display element, and 9 is granule detection zone.
Detailed description of the invention
In order to make the purpose of the present invention, technical scheme and advantage clearly understand, below in conjunction with accompanying drawing, the present invention is further elaborated.
As Figure 1-Figure 2, micro flow control chip device of the present invention, specifically include that DC partial voltage device 4, micro-fluidic chip 5, differential amplification element 6, data acquisition element 7 and display element 8；
As shown in Figure 1, described micro flow control chip device includes intaglio the substrate-PDMS micro-fluidic chip M of some fluid channel, described PDMS micro-fluidic chip M intaglio has fluid channel side and cover plate-glass negative L sealing-in to be structure as a whole, so that forming microchannel between described PDMS micro-fluidic chip and glass negative, substrate-PDMS micro-fluidic chip M material adopts polydimethylsiloxane or poly-methyl methacrylate vinegar, described glass negative L can be substituted by poly-methyl methacrylate vinegar sheet, substrate forms corresponding inlet opening with glass negative by being bonded together after plasma treatment, waste liquid hole, microchannel and detection region；Described microchannel includes main channel 2, focus channel 1, granule detection zone 9 and two detection arm passages 3；
For storing the main channel of analyte sample fluid, these two ends, main channel be respectively arranged to storage detection before sample liquid the first main channel liquid storage hole 21 and for storing the second main channel liquid storage hole 22 of the sample waste after detection；
It is arranged at main channel 2 middle position, the granule detection zone 9 of its detection signal for obtaining and flow through in the analyte sample fluid of one's respective area corresponding to granule, this granule detection zone has two detection arm passages 3, each detection arm passage 3 one end is connected with described main channel, the other end is connected with differential amplification element two input respectively, and this differential amplification element 6 outfan is connected with display element 8 by data acquisition element 7；Each detection arm channel end is respectively arranged with a liquid storage hole for inserting detecting electrode, as imbedded platinum electrode respectively at detection arm channel end；
Play focussing force, for flow in and out two focus channel 1: the first focus channel 11 and the second focus channel 12 that focus on sheath fluid, above-mentioned two focus channel is respectively centered by described granule detection zone, it is symmetricly set in the described granule detection zone left and right sides, before first focus channel 11 one end is arranged to storage detection, the first of ultra-pure water focuses on liquid storage hole 12, and after second focus channel 13 one end is arranged to storage detection, the second of ultra-pure water waste liquid focuses on liquid storage hole 14；
Described first main channel liquid storage hole, the second main channel liquid storage hole, the first focusing liquid storage hole and second focus on liquid storage hole and are connected with DC partial voltage device 4 respectively, this DC partial voltage device for for described first main channel liquid storage hole, the second main channel liquid storage hole, first focus on liquid storage hole and second and focus on liquid storage hole the voltage of different amplitude is provided, to realize testing sample in electric osmose manipulation microchannel and to drive the flowing of ultra-pure water.
Such as Fig. 2, described DC partial voltage device has the output (can apply the voltage of different amplitude) of 4 road different potentials values, thus realizing the flowing of fluid in electric osmose manipulation microchannel, manipulate the flowing of sample solution flowing and ultra-pure water in microchannel including electric osmose；And DC partial voltage device adopts platinum electrode with above-mentioned each liquid storage hole connecting place, to prevent ordinary electrode from being produced cotton-shaped pollutant by electrolysis.
Preferably, upwardly extending bulge-structure 31 is set between described detection arm passage, to form narrower detection zone further so that the ultra-pure water of the sample liquid containing granule to be detected and a focussing force separately flows to each self-corresponding waste liquid hole at narrower region-granule detection zone after meeting again.
The present invention utilizes differential amplification principle that signal is detected, signal is drawn from the sense channel at granule detection zone two ends, thus reaching differential amplification purpose, eliminate common background noise effects, improve signal to noise ratio and accuracy of detection, simultaneously by the first main channel liquid storage hole that is connected with DC partial voltage device respectively, second main channel liquid storage hole, first focuses on liquid storage hole and second focuses on the potential value that liquid storage hole applying is different, make to focus on sheath fluid only to flow in focus channel, to overcome traditional flow focusing method to be sheath fluid from the defect flowing directly into main channel in focus channel.
Described differential amplification element 6 adopts difference amplifier, and its input is connected with the platinum electrode in the liquid storage hole of micro-fluidic chip detection arm channel end by wire.
Specific works process of the present invention is as follows: focus in liquid storage hole all insert platinum electrode at described first main channel liquid storage hole, the second main channel liquid storage hole, the first focusing liquid storage hole and second, the other end of platinum electrode is connected with DC partial voltage device, entered the second main channel liquid storage hole storing waste liquid by granule detection zone in order to driven by electroosmosis analyte sample fluid, driven by electroosmosis ultra-pure water enters the second focusing liquid storage hole of storage waste liquid by granule detection zone simultaneously；By regulating DC partial voltage device so that the potential difference at focus channel two ends, higher than the potential difference at two ends, main channel, namely makes the flow flow higher than sample solution of ultra-pure water；Now in granule detection zone region, ultrapure current can extrude sample liquid so that sample liquid narrowed width；And the Conductivity Ratio sample liquid of ultra-pure water is much larger, it is possible to regard insulant as, therefore the actually detected district of sample liquid narrows, so that detecting device can detect smaller granule.Platinum electrode in the detection arm channel end liquid storage hole at described granule detection zone two ends is connected with the input of difference amplifier each through wire, the outfan of described difference amplifier is connected to the Labview processor (Labview data collecting instrument) for Signal sampling and processing signal, display element adopts PC, in order to realize the detection differential amplification of signal, collection, display and storage.
Detect for little micro-polystyrene sample granule below:
The device parameter of this example: size 30*40*10 μm (width x length × height) in the chip detection region used by the implementation case, detected sample liquid is 3 μm of granules of polystyrene solution；Buffer is phosphate (2 × PBS) buffer solution；The current potential putting on the first main channel liquid storage hole is 18V, and the current potential putting on the second main channel liquid storage hole is 11.5V, and the current potential putting on the first focusing liquid storage hole is 30V, and the current potential putting on the second focusing liquid storage hole is 0V；
Its operating procedure:
(1) in described first main channel liquid storage hole, the second main channel liquid storage hole, add a certain amount of phosphate buffered solution, focus on first in liquid storage hole and the second focusing liquid storage hole and add a certain amount of ultra-pure water；It is inserted into platinum electrode at detection arm channel end liquid storage hole, and it is connected with difference amplifier；Described phosphate buffered solution can configure according to actually required pH value, repeats no more here；
(2) regulating DC partial voltage device makes each output current potential be setting value, and the first corresponding with micro-fluidic chip for each outfan main channel liquid storage hole, the second main channel liquid storage hole, the first focusing liquid storage hole and second are focused on liquid storage hole and be connected；
(3) add a small amount of detected sample liquid at the first main channel liquid storage hole, add same amount of phosphate buffered solution at the second main channel liquid storage hole, focus on first in liquid storage hole and the second focusing liquid storage hole and add same amount of ultra-pure water；
(4) utilizing Labview data collecting instrument and PC, carry out signals collecting, display and storage, its Detection results, such as Fig. 3.
The above; it is only the present invention preferably detailed description of the invention; but protection scope of the present invention is not limited thereto; any those familiar with the art is in the technical scope that the invention discloses; it is equal to replacement according to technical scheme and inventive concept thereof or is changed, all should be encompassed within protection scope of the present invention.
Claims (6)
1. the micro flow control chip device improving resistance impulse method granule accuracy of detection, this device includes intaglio the PDMS micro-fluidic chip of some fluid channel, described PDMS micro-fluidic chip intaglio has fluid channel side and glass negative sealing-in to be structure as a whole, so that forming microchannel between described PDMS micro-fluidic chip and glass negative, it is characterized in that: described microchannel has for the focus channel flowed in and out focusing on sheath fluid, including
For storing the main channel of sample liquid, this main channel has the first main channel liquid storage hole for the sample liquid before storing detection and for storing the second main channel liquid storage hole of the sample waste after detection；
It is arranged at main channel middle position, the granule detection zone of the detection signal for obtaining and flow through in the analyte sample fluid of one's respective area corresponding to granule, this granule detection zone has two one end and is all connected with described main channel, the detection arm passage that the other end is connected with differential amplification element two input respectively, and this differential amplification cell output is connected with display element by data acquisition element；
And focus channel, this focus channel includes two respectively centered by described granule detection zone, it is symmetricly set in the described granule detection zone left and right sides, for realizing the first focus channel and second focus channel of focussing force, and the first focusing liquid storage hole of detection prefocusing sheath fluid it is arranged to store in first focus channel one end, after second focus channel one end is arranged to storage detection, the second of focusing sheath fluid waste liquid focuses on liquid storage hole；
Described first main channel liquid storage hole, the second main channel liquid storage hole, the first focusing liquid storage hole and second focus on liquid storage hole and are connected with DC partial voltage device respectively, this DC partial voltage device for for described first main channel liquid storage hole, the second main channel liquid storage hole, first focus on liquid storage hole and second and focus on liquid storage hole the voltage of different amplitude is provided, to realize in electric osmose manipulation microchannel analyte sample fluid and to drive the flowing focusing on sheath fluid.
2. the micro flow control chip device of raising resistance impulse method granule accuracy of detection according to claim 1, it is characterised in that:
Described first main channel liquid storage hole, the second main channel liquid storage hole, the first focusing liquid storage hole and second focus on liquid storage hole and are connected with DC partial voltage device respectively through platinum electrode.
3. the micro flow control chip device of raising resistance impulse method granule accuracy of detection according to claim 1, it is characterised in that:
Each described detection arm passage is connected with differential amplification element two input respectively through platinum electrode.
4. the micro flow control chip device of raising resistance impulse method granule accuracy of detection according to claim 1, it is characterised in that:
Described data acquisition element adopts Labview data collecting instrument；Described display element adopts PC.
5. the micro flow control chip device of raising resistance impulse method granule accuracy of detection according to claim 1, it is characterised in that:
Described focusing sheath fluid adopts ultra-pure water.
6. the method improving resistance impulse method granule accuracy of detection, it realizes detection by the micro flow control chip device as described in claim 1-5 any one, comprises the steps:
1) in described first main channel liquid storage hole, the second main channel liquid storage hole, add a certain amount of PBS buffer solution, focus on first in liquid storage hole and the second focusing liquid storage hole and add a certain amount of ultra-pure water；In the liquid storage hole of detection arm passage, insert platinum electrode respectively, and aforementioned platinum electrode is connected with differential amplification element respectively；
(2) regulating DC partial voltage device makes each output current potential be setting value, and the first corresponding with micro-fluidic chip for each outfan main channel liquid storage hole, the second main channel liquid storage hole, the first focusing liquid storage hole and second are focused on liquid storage hole and be connected；
(3) add a certain amount of detected sample liquid at the first main channel liquid storage hole, add same amount of phosphate buffered solution at the second main channel liquid storage hole, focus on first in liquid storage hole and the second focusing liquid storage hole and add same amount of ultra-pure water；
(4) utilize Labview data collecting instrument and PC, carry out signals collecting, display and storage.
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| 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 |
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