CN109852542A - A kind of micro-fluidic chip and its processing method for unicellular impedance flow cytometer detection - Google Patents
A kind of micro-fluidic chip and its processing method for unicellular impedance flow cytometer detection Download PDFInfo
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Abstract
A kind of micro-fluidic chip and its processing method for unicellular impedance flow cytometer detection, is related to Cytometric Analysis chip technology field, which includes top layer, electrode-flow channel layer and lower substrate layer.Electrode-flow channel layer includes the runner being made of input section, deferent segment and intermediate detection section and is embedded in intermediate detection section side wall, the differential electrode pair that two pairs of three-diemsnional electrodes contour with side wall of flow channel and being in right opposite placement are constituted.Three-dimensional self-alignment type electrode is bonded without the alignment formula substrate of conventional metals membrane electrode, and detection zone and cell volume to be measured are close to being conducive to improve cell detection sensitivity between the electrode between the three-diemsnional electrode that differential electrode configuration and right opposite are placed.In process aspect, entire electrode-flow channel layer is formed by same material reverse mould, is avoided the additional sacrificial layers photoetching that similar device is generated because using different materials construction electrode and runner, is greatly simplified technique and reduce costs.
Description
Technical field
The present invention relates to Cytometric Analysis chip technology fields, more particularly to a kind of unicellular for no label streaming
The structure of the micro-fluidic chip of detection designs and processing flow.
Background technique
Cell heterogeneity is the universals of biosystem, to the even list of a small number of subgroups in cell colony with heterogeneity
The detection of a cell suffers from important meaning in life science and area of medical diagnostics.Such as to the diseases such as cancer, immune
In the research of disease, often a small number of mutant individuals in group and research object are mostly concerned.Traditional cell detection hand
Duan Duozhen causes the heterogeneity of cell to be ignored the average properties of cell colony, therefore Single cell analysis technology is for detection
The dynamic change of cell colony, disclosing intercellular individual difference is particularly important.
Flow cytometer (flow cytometry), can be by swashing as existing very mature Single cell analysis technology
Light beam carries out the Single cell analysis of fast accurate to the cell of fluorescent marker.Currently based on the miniature fluidic cell of microflow control technique
Instrument reduces costs the miniaturization of this technology and sample consumption.But this technology needs to carry out by agents on cellular outer
Portion's fluorescent marker, it is cumbersome and change cell surface property.In contrast, electrical detection method is examined by label-free mode
The electrical parameter of individual cells is surveyed, easy to operate and cost is lower.Coulter-counter (Coulter Counter) is as most
The representative unicellular electrical detection method of streaming, caused micropore two sides curent change when flowing through micropore by the way that measurement is unicellular
Obtain unicellular volume information, and to develop be medical whole blood instrument.The technical bottleneck of Coulter-counter is only
Suitable for DC electric field cell detection, the cell electrical parameter that can be obtained in addition to cell volume information is extremely limited.And it is based on
Microflow control technique realizes that the unicellular impedance flow cytometer detection technology of multi-frequency receives great attention in recent years.
Unicellular impedance flow cytometer detection technology, which passes through, flows through the cell to suspend in the electrolytic solution successively at a high speed added with fixation
Impedance variations between the electrode and measuring electrode of driving voltage are to obtain inside related cell size, cell membrane and cell membrane
A variety of electrical parameter information.Under low-frequency excitation, to the resistant function of electric current, therefore cell impedance is mainly manifested in cell membrane
The signal amplitude measured in low frequency is positively correlated with cell volume.With the raising of frequency, cell membrane gradually polarizes.In high frequency
Under excitation, cell membrane loses the shielding action to electric current, and cell interior can also be provided in measurement result at this moment in addition to cell volume
Information, such as cytoplasm, nucleus, organelle.
The unicellular streaming impedance detection micro-fluidic chip of early stage is machined with runner and the micro- electricity of metallic film by two panels surface
The substrate alignment of pole is bonded, such as Renaud seminar, Lausanne, SUI university and Southampton University of Southampton Morgan project
Group is invented disclosed in a series of document reports and Chinese patent 201610962524.8 between 2007-2010.It is this kind of
Device needs to be aligned formula, the bonding of the substrate of No leakage, and microelectrode needs to process step by step with fluid channel, and technique is very complicated.
The streaming impedance detection chip reported in the literature for being integrated with the self-alignment type three-diemsnional electrode based on silicon substrate solves to a certain extent
The cumbersome problem of metal film electrode technique of having determined, without being directed at the substep processing of the bonding of formula substrate and electrode, runner, but it is still
It is related to a series of costly and time-consuming silicon technology processes such as dry etching, anode linkage, attenuated polishing.Reported in the literature use is led
Electric polymer, such as silver-dimethyl silicone polymer (AgPDMS), the micro-fluidic chip as self-alignment type three-diemsnional electrode material can be with
It is realized by soft lithographic pour mask technique, greatly reduces the cost and technique of the impedance detection device based on traditional micro fabrication
Complexity.However the prior art reported in the literature is designed using single pair detecting electrode rather than differential electrode, is unfavorable for noise suppressed；
And detecting electrode cross-sectional area and spacing are much larger than cell dia, reduce cell detection sensitivity.Further, since between electrode
Away from larger, which introduces sheath stream focusing structure to stablize position of the cell by electrode when and increase the stability of signal, but
The introducing of sheath stream increases the complexity of design and operation.In terms of technique realization, the prior art is due to being all made of different electricity
Pole (such as AgPDMS) and runner (such as dimethyl silicone polymer, PDMS) material cause chip manufacture to be removed and need process early period
Except mold, the sacrificial layer for defining electrode and flow passage structure step by step, sacrificial layer processing need to also be further processed on mold
Afterwards by electrode filling molding, then sacrificial layer is removed by runner filling molding.And when each making devices, require on mold
Repeat processing sacrificial layer.
A kind of micro-fluidic chip and its processing method for unicellular impedance flow cytometer detection is disclosed herein in the present invention.
Chip utilizes streaming impedance detection technology, and the label-free, electrical parameter of high speed detection cell at different frequencies, it is convenient to operate.
Chip structure is simple, is configured using high sensitivity.The chip manufacture method developed greatly simplifies technique, reduces into
This.
Summary of the invention
In order to overcome above-mentioned defect existing in the prior art, the invention proposes one kind to examine for unicellular impedance streaming
The micro-fluidic chip and its processing method of survey.The self-alignment type three-diemsnional electrode (9) of differential configuration, electrode spacing are used in design
(19) close to unicellular diameter, it ensure that the high sensitivity of chip detection.Side wall of flow channel and electrode are all made of identical conduction
PDMS material construction, can be by low cost and succinct soft light carving technology is realized, the alignment formula without metal film electrode is bonded
Or the dry etch process of silicon based electrode valuableness.The existing chip for being integrated with conductive PDMS electrode is all made of different materials (such as
PDMS runner) is constructed, therefore additional sacrificial layer photoetching need to be carried out on mold during soft lithographic reverse mould and define stream respectively
Road and electrode, runner of the invention and electrode structure are by reverse mould molding of same material without by sacrificial layer photoetching, pole
The processing technology of similar device is simplified greatly.
A kind of micro-fluidic chip for unicellular impedance flow cytometer detection of the invention, the microfluidic chip structure include
It is followed successively by top layer (1), intermediate electrode-flow channel layer (2) and lower substrate layer (3) from top to bottom；
The top layer (1) of chip, which is equipped with, collects the two circular cavities (4) of sample to be tested for being intubated injection and intubation, is respectively aligned to
The injection port (5) and outlet (6) of lower electrode-flow channel layer (2)；
(2) two one side of something of electrode-flow channel layer of chip is respectively equipped with a groove through-hole, is successively denoted as injection port (5) respectively
With outlet (6), centre is equipped with the linear type runner of connection injection port (5) and outlet (6), and runner is divided into three sections: even
Connect the input section (7) of injection port, the deferent segment (8) for connecting outlet, intermediate detection section (10)；
Intermediate detection section (10) two sidewalls are respectively equipped with the groove straight joint perpendicular to linear type runner direction, in groove straight joint
In be also embedded with two pairs of three-diemsnional electrodes (9) composition differential electrode pair；Two electrodes of each pair of three-diemsnional electrode (9) are located at centre
In detection section (10) two sidewalls groove straight joint and just opposite, two opposite electrodes flush in linear type runner with two sidewalls, i.e.,
Electrode height is contour with side wall of flow channel；Two pairs of three-diemsnional electrodes (9) position in intermediate detection section (10) two sidewalls is adjacent and symmetrical；
In the ipsilateral of linear type runner, between non-electrode wall part and three-diemsnional electrode (9), adjacent three-diemsnional electrode is all provided between (9)
Have narrow insulated trenches (11), is filled with insulating materials in insulated trenches (11), is insulated and prevent liquid from insulated trenches
(11) it leaks；
Each electrode of each pair of three-diemsnional electrode (9) includes three sections, from linear type runner intermediate detection section vertically to chip
Electrode-flow channel layer (2) edge be followed successively by the narrow cross-section segments of electrode (12), the electrode changeover portion (13) with tilt angle,
The wide cross-section segments of electrode (14)；The narrow cross-section segments of electrode (12) and the wide cross-section segments of electrode (14) not on the same line, the two
In parallel, electrode changeover portion (13) connects the two, and electrode changeover portion (13) and the narrow cross-section segments of electrode (12) or electrode are wide
The angle α (30) of cross-section segments (14) is at an acute angle.It is ipsilateral in intermediate detection section, the distance between two electrode changeover portions (13) with
It is gradually increased to edge is close；
Electrode and flow passage structure in the electrode-flow channel layer (2) are formed by same conductive PDMS material reverse mould.
Therefore it only needs a reverse mould to can simultaneously obtain the structure of electrode and runner during making devices, avoids the prior art
In since electrode, runner need on mold additional processing sacrificial layer using unlike material respectively define electrode and stream respectively
The complicated technology of road structure.The conductive preferred AgPDMS of PDMS material, it is possible to use carbon-dimethyl silicone polymer (cPDMS) etc..It grinds
Study carefully the result shows that, the AgPDMS for reaching quality score thresholds, which compares cPDMS, higher conductivity, helps to improve the spirit of chip
Sensitivity.
The runner input section (7) and deferent segment (8) preferred 5-5.5mm of length, preferred 50-200 μm of width, runner height
(22) preferably 15-50 μm.It can prevent runner from blocking using wider runner design, and avoid flow resistance excessive.
Preferred 100-500 μm of length of the intermediate detection section (10), intermediate detection section width is equal with electrode spacing (19),
It is preferred that 15-50 μm.Intermediate detection section (10) is conducive to improve the sensitivity of chip detection, shorter inspection using relatively narrow runner design
Survey a possibility that segment length can be to avoid introducing larger flow resistance, reducing channel blockage.
The three-diemsnional electrode width (20), the as length along the direction of linear type runner, preferably 10-30 μm, three-dimensional electricity
High degree (21) is equal with linear type runner height (22).Using the narrow electrode structural designs close with cell dia, cell
(15) when passing through electrode, impedance variations are more apparent, help to improve the sensitivity of chip measurement.Gold is avoided using three-diemsnional electrode
The problem of belonging to membrane electrode alignment.Preferably two pairs of electrode number, electrode placement position is preferably arranged in runner two in the form of face
Side, two electrodes of side apply sinusoidal excitation voltage (16) when connection, and two electrode detection signal of the other side is simultaneously connected with difference form
Line (17) is transferred to subsequent processing circuit, obtains differential signal (18).Differential electrode, which compares single pair electrode, has stronger resist
It makes an uproar ability.The electrode of runner two sides face arrangement compare the ipsilateral arrangement of runner electrode detection signal is more stable and sensitivity more
It is high.
Insulated trenches (11) and non-electrical between the ipsilateral of linear type runner, the narrow cross-section segments of adjacent electrode (12)
Every electrode in pole wall and three-diemsnional electrode (9)-every electrode includes the narrow cross-section segments of electrode (12), has tilt angle
Electrode changeover portion (13), the wide cross-section segments of electrode (14)-between preferred 5-10 μm of width of insulated trenches (11), insulation
Channel is narrower, and capillarity is more obvious.
The highly preferred 2-6mm of the top layer (1)；Quilting material is insulation transparent material, preferably PDMS.The lower substrate layer
(3) insulation transparent material, preferably glass are used.Transparent material facilitates look at experimentation, is also beneficial to the function of extended chip
Can, such as increase fluorescence detection device etc..
A kind of processing method of micro-fluidic chip for unicellular impedance flow cytometer detection of the invention, this method includes mould
Tool processing, electrode-flow channel layer structure formation, top layer pour, demould, punch and be bonded, specifically includes the following steps:
Step (1), mold processing:
A. mask plate is processed according to the pattern that microfluidic chip structure designs；
B., smooth and single-sided polishing silicon wafer (24) is provided, is cleaned up；
C. one layer of SU-8 photoresist of spin coating on silicon wafer (24), thickness are linear type runner height (22)；
D. litho machine and mask plate are used, the silicon wafer (24) for being coated with photoresist is exposed；
E. to the photoresist developing after exposure, the mold (26) with SU-8 photoresist structure (25) is obtained.
The formation of step (2) electrode-flow channel layer structure:
A. PDMS prepolymer and curing agent are mixed with the ratio of mass ratio 10:1, vacuumize process；
B. by the silver powder of 1-3 μm of partial size and PDMS agitation grinding, AgPDMS is made；
C. AgPDMS filler (27) is filled into mold (26) surface, mold (26) overturning is buckled on paper downwards, downwards
Apply pressure, AgPDMS is made to fill up mold (26).It rubs on paper mold (26) surface along vertical electrode direction, grinds off mold
(26) AgPDMS of excess surface keeps AgPDMS filler (27) equal with mold (26) surface；
E. heating solidifies AgPDMS.
Step (3) top layer pours:
A. PDMS prepolymer and curing agent are mixed with the ratio of mass ratio 10:1, vacuumize process after-pouring is in mold
(26) on；
B. heating solidifies PDMS.Cured PDMS can be bonded naturally with lower layer AgPDMS and be integrally formed.
Step (4) demoulding, punching and bonding:
A. PDMS-AgPDMS structure is taken off from mold (26)；
B. hole (4) corresponding with injection port and outlet is processed using punch；
C., the smooth sheet glass of polishing is provided and does lower substrate layer (3), is cleaned up；
D. using the surface AgPDMS of plasma washing machine processing sheet glass upper surface and PDMS-AgPDMS structure；
E. the face AgPDMS is bonded with sheet glass upper surface.
The insulated trenches (11) that following steps block chip can be passed through after the completion of micro Process, comprising: a. is by PDMS prepolymer
It is mixed with curing agent with the ratio of mass ratio 10:1, vacuumize process；B. it takes a small amount of PDMS (28) to be applied to insulated trenches to be located at
At the opening (29) at chip sides edge, so that PDMS is flowed into insulated trenches (11) through capillary action and be heating and curing.
Compared with prior art, advantageous effects of the invention include:
1, runner and electrode structure are made of same conductive PDMS material, and are integrated with conductive PDMS electricity in the prior art
The device of pole is all made of different materials construction runner and electrode, needs to divide on mold by the additional sacrificial layer of lithography process
Not Gou Zao runner and electrode structure, and the mold that the present invention uses can be reused directly once completing the process, without
Sacrificial layer, technique is simplified and cost reduces.
2, runner and electrode structure are made of same conductive PDMS material, with inexpensive and succinct soft lithographic pour mask work
Skill is mutually compatible with；Relative to the existing silicon-based devices for realizing similar function, expensive silicon technology especially dry etching is avoided
Technique.
3, using three-dimensional differential electrode self aligned in design, the existing metallic film electricity for realizing similar function is avoided
Pole device is directed at the bonding of formula substrate.
4, the two pairs of face electrodes connected using difference form, electrode spacing are set using the thin space close with cell dia
Meter is improved compared to the used single pair electrode of integrated electrically conducting PDMS electrode device of similar function and wide detection zone runner design
The sensitivity of detection.
Detailed description of the invention
Fig. 1 is the structural schematic diagram for the micro-fluidic chip of unicellular impedance flow cytometer detection.
Fig. 2 is electrode-runner schematic diagram of a layer structure for the micro-fluidic chip of unicellular impedance flow cytometer detection.
Fig. 3 is the intermediate detection segment structure schematic diagram for the micro-fluidic chip of unicellular impedance flow cytometer detection.
Fig. 4 is the mold structure diagram for the micro-fluidic chip of unicellular impedance flow cytometer detection.
Fig. 5 is the schematic diagram of the process for the micro-fluidic chip of unicellular impedance flow cytometer detection.
Fig. 6 is the filling conduction PDMS material flow diagram for the micro-fluidic chip of unicellular impedance flow cytometer detection；
(a)-(c) sequentially preparation process.
Fig. 7 is the blocking insulated trenches schematic diagram for the micro-fluidic chip of unicellular impedance flow cytometer detection.
Fig. 8 is the scanning electron microscope diagram piece for the micro-fluidic chip of unicellular impedance flow cytometer detection.
Fig. 9 be for unicellular impedance flow cytometer detection micro-fluidic chip under 500kHz and 10MHz frequency to 7 μm, 10 μ
The microballoon of m and 15 μm of partial size measures the high and low frequency amplitude association scatter plot of signal.
Figure 10 be for unicellular impedance flow cytometer detection micro-fluidic chip under 500kHz frequency to 7 μm, 10 μm and 15 μ
The microballoon of m partial size measures the amplitude of signal and microballoon and is associated with scatter plot by the time of differential electrode.
Figure 11 be for unicellular impedance flow cytometer detection micro-fluidic chip under 10MHz frequency to mixing 7 μm with 10 μm
The signal results figure of the microballoon real-time detection of partial size.
Figure 12 is the micro-fluidic chip for unicellular impedance flow cytometer detection simultaneously at 500kHz and 10MHz frequency to mixed
It closes 7 μm of high and low frequency amplitudes for measuring signal with the microballoon of 10 μm of partial sizes and is associated with scatter plot.
Appended drawing reference:
1, top layer；2, electrode-flow channel layer；3, lower substrate layer；4, hole；5, injection port；6, outlet；7, input section；8,
Deferent segment；9, three-diemsnional electrode；10, intermediate detection section；11, insulated trenches；12, the narrow cross-section segments of electrode；13, electrode changeover portion；
14, the wide cross-section segments of electrode；15, cell；16, driving voltage；17, difference form line；18, differential signal；19, between electrode
Away from；20, electrode width；21, electrode height；22, runner height；23, detection zone between electrode；24, silicon wafer；25, SU-8 photoetching
Plastic structure；26, mold；27, AgPDMS filler；28,PDMS；29, insulated trenches are located at the opening at chip sides edge；30,
Angle α.
Specific embodiment
In order to clearly describe objects, features and advantages of the present invention, below in conjunction with the drawings and specific embodiments pair
Design scheme of the invention is described in detail, but the present invention is not restricted to following embodiment.
Embodiment 1
It is specifically shown in Fig. 1-7；
As shown in Figure 1, being the micro-fluidic chip knot for unicellular impedance flow cytometer detection of specific embodiments of the present invention
Structure schematic diagram.
Chip structure includes to be followed successively by PDMS top layer 1, AgPDMS electrode-flow channel layer 2 and glass lower substrate layer from top to bottom
3.1 height of PDMS top layer is 2-6mm, and top layer 1 is used to be intubated injection and 4 diameter of hole of intubation collection sample to be tested is
0.75-2mm。
As shown in Fig. 2, be the micro-fluidic chip for unicellular impedance flow cytometer detection of specific embodiments of the present invention
Electrode-runner schematic diagram of a layer structure.Described 2 liang of one side of something of electrode-flow channel layer are respectively equipped with a groove through-hole, successively remember respectively
For injection port 5 and outlet 6, injection port and outlet diameter are 0.75-3mm, and centre is equipped with a connection injection port 5 and sample out
The linear type runner of mouth 6, runner are divided into three sections, connect the input section 7 of injection port, connect the deferent segment 8 of outlet, intermediate detection
Section 10.Wherein input section 7 and deferent segment 8 take preferred 50-200 μm of larger width to prevent from blocking and introduce larger flow resistance, and
Equal length, preferably 5-5.5mm.
10 two sidewalls of intermediate detection section are respectively equipped with the groove straight joint perpendicular to linear type runner direction, in groove straight joint
Also it is embedded with the differential electrode pair of two pairs of three-diemsnional electrodes 9 composition；Two electrodes of each pair of three-diemsnional electrode 9 are located at intermediate detection section
In 10 two sidewalls groove straight joints and just opposite, two opposite electrodes flush in linear type runner with two sidewalls, i.e. electrode height
It is contour with side wall of flow channel；Two pairs of three-diemsnional electrodes 9 position in 10 two sidewalls of intermediate detection section is adjacent and symmetrical；Each pair of three-diemsnional electrode
9 each electrode includes three sections, from linear type runner intermediate detection section vertically to the edge of electrode-flow channel layer 2 of chip according to
Secondary is the narrow cross-section segments 12 of electrode, the wide cross-section segments 14 of electrode changeover portion 13, electrode with tilt angle；The narrow cross section of electrode
Not on the same line with the wide cross-section segments 14 of electrode, the two is parallel, and electrode changeover portion 13 connects the two, electrode for section 12
Changeover portion 13 and the angle α 30 of the narrow cross-section segments 12 of electrode or the wide cross-section segments 14 of electrode are at an acute angle.It is ipsilateral in intermediate detection section,
The distance between two electrode changeover portions 13 are gradually increased with to edge is close.Extend to the wide cross-section segments 14 of chip edge electrode
Preferred 100-400 μm of width.Overall length preferred 2-5mm of the electrode from test side to chip edge.Ipsilateral interelectrode insulated trenches
11 guarantee that cell successively passes through two pairs of differential electrode clock synchronization local environments using preferred 5-10 μm of relatively narrow width in detection zone
Consistency, the insulated trenches move towards width with electrode during extending to chip edge and accordingly become larger.Electrode and non-electrode wall
Preferred 5-10 μm of 11 width of insulated trenches between body portion.It is filled with insulating materials in insulated trenches 11, insulated and is prevented
Only liquid is leaked from insulated trenches 11.
As shown in figure 3, be the micro-fluidic chip for unicellular impedance flow cytometer detection of specific embodiments of the present invention
Detect segment structure schematic diagram.10 width of intermediate detection section is equal with electrode spacing 19 and close to surveyed cell dia size, thus
It ensure that the sensitivity of cell detection.Embodiment simulates cell 15 using 7-15 μ m diameter microballoon, therefore it is wide to detect section runner
Degree should be slightly bigger than 15 μm, preferred 100-500 μm of length.Runner height 22, i.e., described electrode-flow channel layer thickness, preferably 15-50 μ
m.The three-diemsnional electrode 9 being embedded in the groove straight joint of 10 two sidewalls of intermediate detection section, electrode height 21 and 22 phase of runner height
Deng, close the surveyed cell dia size of width 20, slightly larger than 15 μm, as shown in Figure 8.According to this design, electrode positive area with
Spacing minimizes under the premise of allowing cell to be measured to pass through, so that 23 volume of detection zone is close between the electrode of face
15 volume of cell, make by cell by bring impedance variations it is more obvious, improve chip detection sensitivity and reduce due to
The difference bring signal intensity that cell passes through position when electrode.
As shown in figure 4, being the micro-fluidic chip mould for unicellular impedance flow cytometer detection of specific embodiments of the present invention
It signs an undertaking composition.The existing chip for being integrated with AgPDMS electrode is all made of different materials construction runner, such as PDMS.Add man-hour requirement
Carry out sacrificial layer photoetching additionally on mold to construct electrode and flow passage structure respectively.Specifically, passing through mold and sacrificial layer figure
Case common definition electrode removes sacrificial layer after the completion of the filling of AgPDMS electrode, and PDMS stream is filled at original sacrifice layer pattern
Road structure.The technique requires to re-work sacrificial layer on mold in each reverse mould device.Electrode-flow channel layer of the invention
2 are made of same material AgPDMS, and 26 pattern of mold defines electrode and flow passage structure simultaneously, are not necessarily to additional processing sacrifice
Layer.Mold 26 can be reused directly once completing the process, and greatly simplify the processing technology of similar device.
As shown in figure 5, the micro-fluidic chip for unicellular impedance flow cytometer detection for specific embodiments of the present invention adds
Work flow chart.The processing of chip needs to carry out in ultra-clean chamber, including mold processing, electrode-flow channel layer structure formation, top layer pour
It builds, demould, punch and is bonded.
The processing of step (1) mold:
A. mask plate is processed according to the pattern that microfluidic chip structure designs；
B., smooth and single-sided polishing 4 inch silicon wafer 24 is provided, is cleaned up；
C. one layer of SU-8 photoresist of spin coating on silicon wafer 24, preferred 15-50 μm of thickness；
D. litho machine and mask plate are used, the silicon wafer 24 for being coated with photoresist is exposed.
E. to the photoresist developing after exposure, the mold 26 with SU-8 photoresist structure 25 is obtained.Step (2) electrode-
The formation of flow channel layer structure:
A. PDMS prepolymer and curing agent are mixed and stirred for uniformly with the ratio of mass ratio 10:1,
B. the silver powder that partial size is 1-3 μm is uniformly mixed again with aforementioned PDMS mixture and AgPDMS is made, wherein silver powder accounts for
The 85% of AgPDMS mixture quality；
C. as shown in Fig. 6 (a), AgPDMS filler 27 is filled into 26 surface of mold；As shown in Fig. 6 (b), then by mold 26
Overturning is buckled on paper downwards, is downwardly applied to pressure, AgPDMS is made to fill up mold 26, is rubbed on paper mould along vertical electrode direction
Have 26 surfaces, grind off the AgPDMS of 26 excess surface of mold, keeps AgPDMS filler 27 equal with 26 surface of mold, such as Fig. 6 (c)
It is shown；
D. the placement of mold 26 that will be filled with AgPDMS is heating and curing on hot plate, and preferred 60-95 degrees Celsius of temperature, baking
It is preferred that 1-3 hours.
Step (3) top layer pours:
A. PDMS prepolymer and curing agent are mixed and stirred for uniformly with the ratio of mass ratio 10:1, vacuumize process；
B. aluminium-foil paper is folded into the box-like of no ceiling, box is slightly larger than mold 26, by mold 26 as aluminium foil carton
It is interior, PDMS mixture is cast in 26 surface of mold；
C. the aluminium foil carton for holding the mold 26 for having poured PDMS is purchased by oneself in the PDMS that is heating and curing in heating plate, temperature is excellent
60-95 degrees Celsius is selected, is toasted 1-3 hours preferred.
Step (4) demoulding, punching and bonding:
A. it will solidify and the PDMS-AgPDMS structure of bonding one taken off from mold 26 naturally；
B. hole corresponding with injection port and outlet is processed using punch, cleans the structure；
C., the smooth sheet glass of polishing is provided and does lower substrate layer 3, cleaning base plate；
D. using the surface AgPDMS about 60 of plasma washing machine processing sheet glass upper surface and PDMS-AgPDMS structure
Second；
E. the face AgPDMS is bonded with sheet glass upper surface.
As shown in fig. 7, the micro-fluidic chip for unicellular impedance flow cytometer detection for specific embodiments of the present invention is stifled
Fill in 11 schematic diagram of insulated trenches.
A. PDMS prepolymer and curing agent are sufficiently mixed with the ratio of mass ratio 10:1, vacuumize process；
B. a small amount of PDMS28 of picking is applied to insulated trenches and is located at the opening 29 at chip sides edge, keeps PDMS28 logical
It crosses capillarity and flows into insulated trenches 11 to block insulated trenches, then chip is placed in 120 degree of heating plates and toasts 10 points
Clock solidifies rapidly PDMS.
Fig. 2 goes back while embodying the micro-fluidic chip for unicellular impedance flow cytometer detection of specific embodiments of the present invention
Connection configures, and two pairs of face three-diemsnional electrodes 9 are connected in this example with difference form.Electrode in the side of chip linear type runner is applied
Add same driving voltage 16, which is the superposition of the equal sinusoidal voltage of the amplitude of one or more frequencies, the other side
Two electrodes receive signal with difference form line 17, and input subsequent amplification circuitry.When it is cell-free by when, between two pairs of electrodes
Impedance differences be zero.When having cell 15 by changing this when any pair of electrode to interelectrode impedance, to generate difference
Sub-signal 18.Differential electrode compares single pair electrode with stronger anti-noise ability.The electrode of runner two sides face arrangement compares stream
The electrode of the ipsilateral arrangement in road, more evenly, detection signal is more stable and sensitivity is higher for field distribution.
Polystyrene microsphere is used to carry out impedance analysis test to the chip as cell model, at multiple frequencies together
When collect impedance test signal.Specific embodiment includes:
Step 1, connection chip and signal processing equipment equipment:
Chip microelectrode and the transmission of the electric signal line of follow-up signal process instrumentation equipment use following methods: in three-dimensional electricity
The end profile that pole 9 is located at the wide cross-section segments 14 of each electrode of chip edge smears elargol, and thin by elargol and one
Soft copper conducting wire one end is connected, and conducting wire other end is welded on pcb board and is transferred to SMA interface by being routed on pcb board.Chip is straight
A pair of of three-diemsnional electrode of line style runner the same side is by above-mentioned bus connection method by the SMA interface on pcb board come self-impedance point
Analyzer motivates the driving voltage 16 of output end, and a pair of of three-diemsnional electrode of chip linear type runner other side passes through above-mentioned line side
Method is connected to multi-frequency impedance analyzer by a preamplifier with difference form line 17 by the SMA interface on pcb board
Input terminal simultaneously carries out subsequent processing.
Step 2 prepares sample solution:
It configures phosphate buffer (PBS).The clean glass reagent bottle that capacity is 1L is taken, PBS solution proportion is phosphoric acid
Above-mentioned solute is dissolved in 1L deionized water by potassium dihydrogen 0.27g, disodium hydrogen phosphate 1.42g, sodium chloride 8g, potassium chloride 0.2g,
It is uniformly mixed, obtains PBS solution, conductivity 1.5S/m.Microsphere sample is the preparation method is as follows: taking concentration is about 5 × 107A/ml
7 μm of microspheres solutions, 10 μ l, with 990 μ l phosphate buffer injection 1ml centrifuge tube be uniformly mixed, be made concentration be 5 × 105
7 μm of microsphere samples solution of a/ml；Taking concentration is about 2 × 10710 μm of microspheres solution 10ul of a/ml, with the phosphoric acid of 990 μ l
Salt buffer injects 1ml centrifuge tube and is uniformly mixed, and it is 2 × 10 that concentration, which is made,510 μm of microsphere samples solution of a/ml；Take concentration
About 5 × 10615 μm of microspheres solution 20ul of a/ml, the phosphate buffer injection 1ml centrifuge tube with 980 μ l are uniformly mixed,
It is 1 × 10 that concentration, which is made,515 μm of microsphere samples solution of a/ml；Taking concentration is about 5 × 1077 μm of 10 μ of microspheres solution of a/ml
L, taking concentration is about 2 × 10710 μm of microspheres solution 10ul of a/ml, the phosphate buffer with 980 μ l inject 1ml centrifuge tube
It is uniformly mixed, 7 μm and 10 μm of microballoon mixing sample solution is made.
Step 3, fluid driving equipment:
Microsphere samples solution to be measured is pumped into the hole 4 that chip top-layer is directed at injection port 5 by pressure control equipment,
Setting driving pressure is 2mbar.By stainless between chip top-layer hole 4 and the sample output channel of pressure control equipment driving
Steel pipe connection, stainless steel tube diameter is slightly larger than the sample output channel that chip top-layer hole 4 and pressure control equipment drive.It is setting
It sets under pressure, microballoon is by the speed of electrode up to 2.5mm/s.
Step 4, impedance signal acquisition:
The excitation output of impedance analyzer is set, and opening pressure driving makes microballoon by electrode, opens simultaneously impedance analysis
The excitation of instrument switchs and collects detection data.
Step 5, bifrequency real-time impedance signal measurement:
The superposed signal that impedance analyzer excitation output end output includes high and low two kinds of frequency contents is set, respectively
500kHz, 0.625Vpp and 10MHz, 0.625Vpp.
To above-mentioned 7 μm, 10 μm, 15 μm, 7 μm of mixing and 10 μm of four kinds of microsphere samples solution, respectively repeat steps 3-4 into
Row test, is not less than 200 by the microballoon quantity of electrode when every group of test sample.
Test result is as follows:
1, for the microballoon of same partial size, by taking the test result under step 5 setting as an example, microspherulite diameter size is in low frequency
With the primary factor under high frequency being influence detection signal amplitude, i.e. partial size is bigger, and the impedance signal measured is more obvious.High frequency
Detection signal amplitude under (10MHz) is all larger than the detection signal amplitude under low frequency (500kHz), this be low frequency detection signal by
Caused by the influence that microelectrode electric double layer effect divides driving voltage.What low frequency signal mainly reflected is microspherulite diameter size institute
Inhibition of the microsphere surface electric double layer of decision to signal.The influence of the electric double layer of microelectrode and microsphere surface disappears under high frequency
It loses, measures signal while being decided by the electrical parameter (conductivity, dielectric constant etc.) of microspherulite diameter size and microballoon.Test result
The high and low frequency signal proportion for showing every kind of partial size microballoon is definite value, this is because three kinds of microballoons are polystyrene material, tool
Have an identical electrical parameter, and due to microspherulite diameter, and bring signal difference different by position when electrode by high frequency with
Low frequency signal is done than excluding, therefore measures identical low-and high-frequency signal ratio, as shown in Figure 9.
2, analyze 7 μm, 10 μm, 15 μm of three kinds of microsphere samples solution respectively step 5 setting under test result, microballoon warp
The time for crossing two pairs of three-diemsnional electrodes is elongated with microspherulite diameter increase.This is because in the chip of identical electrodes spacing, due to
Pressure-driven fluid it is existing be parabola property velocity flow profile, i.e., closer to side wall of flow channel, flow velocity is smaller.Since partial size is larger
Its boundary of microballoon microballoon more lesser than partial size closer to electrode sidewall, drive the fluid flow rate of microballoon movement also due to leaning on nearside
Wall and it is smaller, cause microballoon by pole time it is longer, as shown in Figure 10.
3, it is tested under the setting of step 5 with 10 μm of mixing microsphere samples solution for 7 μm, resulting real-time detection letter
Number can clearly by signal amplitude distinguish two kinds of microballoons, as shown in figure 11；It can also be obtained simultaneously by statistical test signal
The amplitude scatter plot that the longitudinal axis is high frequency, horizontal axis is low frequency, scatter plot can clearly distinguish two scatterplot aggregation clusters, respectively
Two kinds of microsphere components in mixing sample are represented, as shown in figure 12.
Claims (9)
1. a kind of micro-fluidic chip for unicellular impedance flow cytometer detection, which is characterized in that the microfluidic chip structure packet
Containing being followed successively by top layer (1), intermediate electrode-flow channel layer (2) and lower substrate layer (3) from top to bottom；
The top layer (1) of chip, which is equipped with, collects the two circular cavities (4) of sample to be tested for being intubated injection and intubation, is respectively aligned to lower layer
The injection port (5) and outlet (6) of electrode-flow channel layer (2)；
(2) two one side of something of electrode-flow channel layer of chip is respectively equipped with a groove through-hole, is successively denoted as injection port (5) respectively and goes out
Sample mouth (6), centre are equipped with the linear type runner of connection injection port (5) and outlet (6), and runner is divided into three sections: connect into
The input section (7) of sample mouth, the deferent segment (8) for connecting outlet, intermediate detection section (10)；
Intermediate detection section (10) two sidewalls are respectively equipped with the groove straight joint perpendicular to linear type runner direction, in groove straight joint also
It is embedded with the differential electrode pair of two pairs of three-diemsnional electrodes (9) composition；Two electrodes of each pair of three-diemsnional electrode (9) are located at intermediate detection
In the groove straight joint of section (10) two sidewalls and just opposite, two opposite electrodes flush in linear type runner with two sidewalls, i.e. electrode
Height is contour with side wall of flow channel；Two pairs of three-diemsnional electrodes (9) position in intermediate detection section (10) two sidewalls is adjacent and symmetrical；Straight
Line style runner it is ipsilateral, be equipped between non-electrode wall part and three-diemsnional electrode (9), between adjacent three-diemsnional electrode (9) narrow
Narrow insulated trenches (11), insulated trenches (11) are interior to be filled with insulating materials, is insulated and prevents liquid from insulated trenches (11)
Leakage.
2. a kind of micro-fluidic chip for unicellular impedance flow cytometer detection described in accordance with the claim 1, which is characterized in that every
Each electrode to three-diemsnional electrode (9) includes three sections, from linear type runner intermediate detection section vertically to the electricity of chip
The edge of pole-flow channel layer (2) is followed successively by the narrow cross-section segments of electrode (12), electrode changeover portion (13), electrode with tilt angle
Wide cross-section segments (14)；Not on the same line, the two is parallel for the narrow cross-section segments of electrode (12) and the wide cross-section segments of electrode (14),
Electrode changeover portion (13) with tilt angle connects the two, electrode changeover portion (13) and electrode with tilt angle
The angle α (30) of narrow cross-section segments (12) or the wide cross-section segments of electrode (14) is at an acute angle；It is ipsilateral in intermediate detection section, two electrodes tool
There is the distance between electrode changeover portion (13) of tilt angle to be gradually increased with to edge is close.
3. a kind of micro-fluidic chip for unicellular impedance flow cytometer detection described in accordance with the claim 1, which is characterized in that institute
State runner input section (7) and deferent segment (8) preferred 5-5.5mm of length, preferred 50-200 μm of width, linear type runner height (22)
It is preferred that 15-50 μm；
Preferred 100-500 μm of length of the intermediate detection section (10), intermediate detection section width is equal with electrode spacing (19), preferably
15-50μm；
The three-diemsnional electrode width (20), the as length along the direction of linear type runner, preferably 10-30 μm, three-diemsnional electrode are high
It is equal with linear type runner height (22) to spend (21).
4. a kind of micro-fluidic chip for unicellular impedance flow cytometer detection described in accordance with the claim 1, which is characterized in that electricity
Pole placement position is preferably arranged in runner two sides in the form of face, and two electrodes of side apply driving voltage (16) when connection, should
Driving voltage is the superposition of one or more different frequencies and the sinusoidal voltage of identical amplitude, and two electrode detection signal of the other side is simultaneously
Subsequent amplifying circuit is transferred to difference form line (17)；When it is cell-free by when, the impedance between two pairs of electrodes is equal,
Output difference signal is zero；When there is cell (15) successively to pass through two pairs of electrodes, can change between the electrode pair passed through
Impedance, to generate differential signal (18).
5. a kind of micro-fluidic chip for unicellular impedance flow cytometer detection according to claim 2, which is characterized in that phase
Insulated trenches (11) and the narrow cross-section segments of electrode (12), the electricity with tilt angle between the adjacent narrow cross-section segments of electrode (12)
The preferred 5-10 μ of width of insulated trenches (11) between pole changeover portion (13), the wide cross-section segments of electrode (14) and non-electrode wall
m。
6. a kind of micro-fluidic chip for unicellular impedance flow cytometer detection described in accordance with the claim 1, which is characterized in that institute
Electrode flow channel layer is stated to be formed by same conductive dimethyl silicone polymer (conductive PDMS) material reverse mould；Conductive PDMS material is preferred
Silver-colored dimethyl silicone polymer (AgPDMS) or carbon poly dimethyl siloxane (cPDMS)；During making devices, electrode and runner
Structure only needs a reverse mould that can form simultaneously.
7. a kind of micro-fluidic chip for unicellular impedance flow cytometer detection described in accordance with the claim 1, which is characterized in that institute
State top layer (1) highly preferred 2-6mm；Quilting material is insulation transparent material, preferably dimethyl silicone polymer (PDMS)；Under described
Substrate layer (3) uses insulation transparent material, preferably glass.
8. a kind of described in any item processing sides of the micro-fluidic chip for unicellular impedance flow cytometer detection claim 1-7
Method, this method include mold processing, and electrode-flow channel layer structure formation, top layer pours, demoulds, punches and be bonded, specific to wrap
It includes:
The processing of step (1) mold:
A. mask plate is processed according to the pattern that microfluidic chip structure designs；
B., smooth and single-sided polishing silicon wafer (24) is provided, is cleaned up；
C. one layer of SU-8 photoresist of spin coating on silicon wafer (24), thickness are linear type runner height (22)；
D. litho machine and mask plate are used, the silicon wafer (24) for being coated with photoresist is exposed；
E. to the photoresist developing after exposure, the mold (26) with SU-8 photoresist structure (25) is obtained；
The formation of step (2) electrode-flow channel layer structure:
A. PDMS prepolymer and curing agent are mixed with the ratio of mass ratio 10:1, vacuumize process；
B. by the silver powder of 1-3 μm of partial size and PDMS agitation grinding, AgPDMS is made；
C. AgPDMS filler (27) is filled into mold (26) surface, mold (26) overturning is buckled on paper downwards, is downwardly applied to
Pressure makes AgPDMS fill up mold (26)；It rubs on paper mold (26) surface along vertical electrode direction, grinds off mold (26)
The AgPDMS of excess surface keeps AgPDMS filler (27) equal with mold (26) surface；
D. heating solidifies AgPDMS；
Step (3) top layer pours:
A. PDMS prepolymer and curing agent are mixed with the ratio of mass ratio 10:1, vacuumize process after-pouring is in mold (26)
On；
B. heating solidifies PDMS；Cured PDMS can be bonded naturally with lower layer AgPDMS and be integrally formed；
Step (4) demoulding, punching and bonding:
A. PDMS-AgPDMS structure is taken off from mold (26)；
B. hole (4) corresponding with injection port and outlet is processed using punch；
C., the smooth sheet glass of polishing is provided and does lower substrate layer (3), is cleaned up；
D. using the surface AgPDMS of plasma washing machine processing sheet glass upper surface and PDMS-AgPDMS structure；
E. the face AgPDMS is bonded with sheet glass upper surface.
9. a kind of processing method of the micro-fluidic chip for unicellular impedance flow cytometer detection according to claim 8,
It is characterized in that, insulated trenches (11) are blocked after the completion of micro Process, and step includes by PDMS prepolymer and curing agent with matter
Measure the ratio mixing than 10:1, vacuumize process；It takes a small amount of PDMS (28) to be applied to insulated trenches and is located at chip sides edge
It is open at (29), so that PDMS is flowed into insulated trenches (11) through capillary action and be heating and curing.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110711608A (en) * | 2019-09-23 | 2020-01-21 | 中国科学院上海微系统与信息技术研究所 | Microfluidic chip for cell detection and preparation method thereof |
| CN111272835A (en) * | 2020-03-27 | 2020-06-12 | 大连海事大学 | Non-contact capacitance detection device and method for oil pollutants |
| CN113155688A (en) * | 2021-05-18 | 2021-07-23 | 南京智能高端装备产业研究院有限公司 | Multifunctional electrical detection chip for detecting cell parameters |
| CN113804735A (en) * | 2021-09-01 | 2021-12-17 | 西北工业大学 | High-performance single-cell impedance detection microelectrode and preparation method thereof |
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| CN114295688A (en) * | 2021-12-30 | 2022-04-08 | 北京航空航天大学 | Electrical impedance tomography sensor suitable for microchannel heterogeneous object detection |
| CN114870917A (en) * | 2022-05-09 | 2022-08-09 | 南京大学 | Microfluidic chip for identifying different cells and preparation method and detection platform thereof |
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7250775B1 (en) * | 2003-11-12 | 2007-07-31 | The Regents Of The University Of California | Microfluidic devices and methods based on measurements of electrical admittance |
| CN102460114A (en) * | 2009-06-05 | 2012-05-16 | 皇家飞利浦电子股份有限公司 | Multi -frequency impedance method and apparatus for discriminating and counting particles expressing a specific marker |
| WO2014170625A1 (en) * | 2013-04-16 | 2014-10-23 | University Of Southampton | Apparatus for electrically measuring individual particles flowing in a liquid |
| CN106492891A (en) * | 2016-10-28 | 2017-03-15 | 中国人民解放军军事医学科学院卫生装备研究所 | Electrical impedance flow cytometer detection molecule, the micro-fluidic chip of cell and preparation method |
| WO2017106727A1 (en) * | 2015-12-16 | 2017-06-22 | President And Fellows Of Harvard College | Electrode integration into organs on chip devices |
| CN206577776U (en) * | 2016-10-28 | 2017-10-24 | 中国人民解放军军事医学科学院卫生装备研究所 | Electrical impedance flow cytometer detection molecule, the micro-fluidic chip of cell |
| CN108507910A (en) * | 2018-03-16 | 2018-09-07 | 中国科学院大学 | A kind of microfluidic chip devices of detection Atmospheric particulates |
-
2018
- 2018-12-18 CN CN201811555345.8A patent/CN109852542B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7250775B1 (en) * | 2003-11-12 | 2007-07-31 | The Regents Of The University Of California | Microfluidic devices and methods based on measurements of electrical admittance |
| CN102460114A (en) * | 2009-06-05 | 2012-05-16 | 皇家飞利浦电子股份有限公司 | Multi -frequency impedance method and apparatus for discriminating and counting particles expressing a specific marker |
| WO2014170625A1 (en) * | 2013-04-16 | 2014-10-23 | University Of Southampton | Apparatus for electrically measuring individual particles flowing in a liquid |
| WO2017106727A1 (en) * | 2015-12-16 | 2017-06-22 | President And Fellows Of Harvard College | Electrode integration into organs on chip devices |
| CN106492891A (en) * | 2016-10-28 | 2017-03-15 | 中国人民解放军军事医学科学院卫生装备研究所 | Electrical impedance flow cytometer detection molecule, the micro-fluidic chip of cell and preparation method |
| CN206577776U (en) * | 2016-10-28 | 2017-10-24 | 中国人民解放军军事医学科学院卫生装备研究所 | Electrical impedance flow cytometer detection molecule, the micro-fluidic chip of cell |
| CN108507910A (en) * | 2018-03-16 | 2018-09-07 | 中国科学院大学 | A kind of microfluidic chip devices of detection Atmospheric particulates |
Non-Patent Citations (4)
| Title |
|---|
| BIANCHI E, ET AL.: "Detecting particles flowing through interdigitated 3D microelectrodes.", 《CONF PROC IEEE ENG MED BIOL SOC.》 * |
| DE NINNO A, ET AL.: "Coplanar electrode microfluidic chip enabling accurate sheathless impedance cytometry.", 《LAB ON A CHIP》 * |
| LEWPIRIYAWONG N, ET AL.: "AC-dielectrophoretic characterization and separation of submicron and micron particles using sidewall AgPDMS electrodes.", 《BIOMICROFLUIDICS.》 * |
| 唐文来，等: "基于微流控技术的单细胞生物物理特性表征", 《化学进展》 * |
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110711608A (en) * | 2019-09-23 | 2020-01-21 | 中国科学院上海微系统与信息技术研究所 | Microfluidic chip for cell detection and preparation method thereof |
| CN110711608B (en) * | 2019-09-23 | 2022-03-15 | 中国科学院上海微系统与信息技术研究所 | Microfluidic chip for cell detection and preparation method thereof |
| CN111272835A (en) * | 2020-03-27 | 2020-06-12 | 大连海事大学 | Non-contact capacitance detection device and method for oil pollutants |
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| CN113804735A (en) * | 2021-09-01 | 2021-12-17 | 西北工业大学 | High-performance single-cell impedance detection microelectrode and preparation method thereof |
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| CN114295688A (en) * | 2021-12-30 | 2022-04-08 | 北京航空航天大学 | Electrical impedance tomography sensor suitable for microchannel heterogeneous object detection |
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| CN115126931B (en) * | 2022-06-08 | 2023-08-11 | 厦门大学 | Microfluidic chip, manufacturing method thereof and electromagnetic control valve device |
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| CN115319434B (en) * | 2022-09-15 | 2024-03-19 | 中国矿业大学 | Automatic clamping and intubation device and method for microfluidic chip |
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