CN208060350U - A kind of micro flow control chip device particle being carried out based on nano-micrometre combination of channels and cell sequence is detached and counted - Google Patents
A kind of micro flow control chip device particle being carried out based on nano-micrometre combination of channels and cell sequence is detached and counted Download PDFInfo
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- CN208060350U CN208060350U CN201820526713.5U CN201820526713U CN208060350U CN 208060350 U CN208060350 U CN 208060350U CN 201820526713 U CN201820526713 U CN 201820526713U CN 208060350 U CN208060350 U CN 208060350U
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Abstract
The utility model discloses a kind of micro flow control chip devices carrying out particle based on nano-micrometre combination of channels and cell sequence is detached and counted, and microchannel includes：Sample intake passage；Main channel；Multiple sample output passages；There is first electric field separates counting channel separation to count positive liquid storage hole, and pass through the first detection door and main channel vertical connection；There is second electric field separates counting channel separation to count cathode liquid storage hole, and pass through the second detection door and main channel vertical connection and flow field focus channel；Separation counts in positive/negative liquid storage hole and is inserted with platinum electrode, and DC power anode is connect with the first electric field separates counting channel, and cathode is connect with the second electric field separates counting channel；DC power supply cascode reference resistance is simultaneously connect by it with signal amplification component, and then is connect with signal acquisition control system；The utility model enables to cell to be detected to realize counting without entering detection door and under conditions of realization does not destroy cell to be separated, different sample output passages is accurately flowed under dielectrophoretic force effect.
Description
Technical field
The utility model is related to particle monitoring technique fields, particularly relate to a kind of based on nano-micrometre combination of channels
It carries out particle and cell sequence detaches and the micro flow control chip device of counting.
Background technology
Not allogenic cell in mixing sample is detached, and the cell after separation is accurately counted in multiple fields
Have great importance and demand；For example, in marine environmental monitoring, the fields such as biomedical research are mixed to determining target
Close the dress that the portable cell quick separating of the quantity of object (such as bacterium, virus, marine microorganism) variety classes cell counts
It sets or method, there is urgent demand always.
Carrying out DEP separation in micro-fluidic field has direct current dielectrophoresis and AC dielectric swimming two types.However it is situated between in direct current
In order to generate the voltages that the dielectrophoretic force of sufficient intensity generally requires the even upper kilovolt of hundreds of volts in electrophoresis, and what high voltage was formed
Electric field will produce joule heating effect again, and cell or microorganism are easy to be killed or crack in this environment；It is situated between for exchange
For electrophoresis, it usually needs complicated microelectrode array or microelectrode structure is aligned with microfluidic channel with high accuracy, it is this
Operation is difficult to realize.
For RPS detections, particle need to enter detection door when traditional RPS is detected, and will appear particle will detect door
The case where blocking, this can make testing result mistake occur, and accuracy of detection substantially reduces reduction.
Additionally due to tradition DEP separation needs high voltage, when voltage is very high, detect that the signal-to-noise ratio of signal can increase therewith
Greatly；With the increase of terminal voltage, the noise of system can become larger, so excessively high voltage can generate shadow to the result of grain count
It rings.
The method of common cell separation existing at present includes following several：
1) it is separated by filtration method：The method of being separated by filtration is to detach the even common method of item mixture at present, it is one kind by more
The obstacle ground that hole medium is constituted makes the method that various sizes of cell is detached.The method principle is simple, but requires machining for
The very high porous media of precision (such as filter membrane, strainer), and filtering accuracy is bad, cannot detach cell similar in two kinds of volumes.
2) density gradient ultracentrifugation rate partition method：It is it is a kind of using colloidal solid under centrifugal field force effect pass through it is close
The rate for spending gradient zones is different, by controlling parameter of noncentricity, realizes nano particle according to differences such as size, density and aggregating states
The method detached.The method detaches particle using the density and size of particle, but in ultracentrifugal physics item
Under part, some cells can be destroyed, and separable molecule has certain limitation.
3) field flow classification method：It is a kind of important means of particle separation, and field flow classification method is in a long and narrow tunnel
In road, field is applied into suspension or solution therein, is detached in the direction of mobile phase with vertical (or other angles)
Method, this method can the colloid wider to size range continue high-resolution separation, but this method deficiency
It being in and is easily lost in sample, main cause is during sample and seperation film act on, and seperation film can generate absorption to sample,
PH value, ionic strength of sample etc. can also change.
4) Magnet Treatment：Magnet Treatment is a kind of method being effectively separated to cell using magnetic action power,
Equipment volume needed for this method is small, simple in structure, maintenance is easy, but sometimes for raising magnetic field intensity, it is necessary to selection is high
The magnetic matrix of magnetic saturation has certain technical difficulty to the selection of magnetic matrix, and increases the expense of operation.
Micro-fluidic chip is built on as a kind of new micro-analysis platform in the 1990s, it passes through microfabrication skill
Art builds the microflow path system being made of micro- function element such as liquid storage tank, micro- reative cell, microchannels on chip, loads biological sample
After product and reaction solution, microfluidic circuit is formed under pressure or electric field action, with chip carry out it is a kind of or it is continuous it is a variety of react, reach
To the purpose that sample is quickly analyzed, micro fluidic device can by biology, chemistry, the sample preparation of medical analysis process, reaction, point
It is integrated on the chip of one piece of micro-meter scale from basic operations such as detections, is automatically performed analysis overall process.It has liquid flow
It is dynamic controllable, consumption reagent and sample are few, the features such as improving to analyze speed tenfold hundreds of times.
Initial separating chips structure include a wide main channel of hundreds of microns and setting with main channel middle part and it is vertical
Electric field channel in main channel is connect using platinum electrode with power supply at electric field channel both ends, since electric field channel and main channel are handed over
Electric field strength very little at remittance needs prodigious voltage, when cell volume very little is needed electricity to obtain good separating effect
Press very high, this is difficult to realize；The intersection of electric field channel and main channel is made several microns very by later separating chips
To the flow field focus channel for being Nano grade, this keeps electric field strength very big in intersection, to make voltage needed for separation reduce；Most
New separating chips add a channel again below sample intake passage, and PBS buffer solution is added in the channel, cell is made to exist
The one side wall flowing that main channel is pasted under the action of buffer solution pressure, to which by the highest point of electric field strength on main channel, this makes
The cell dielectrophoretic force that place is received on one point increases, and substantially reduces voltage needed for separation.
Further, since split tunnel and counting channel are closer to, it is easy to form interference mutually, influence the movement of particle
With the order of accuarcy of counting.
Invention content
In view of defects in the prior art, the purpose of this utility model is to provide for a kind of based on nano-micrometre channel
The micro flow control chip device that combination carries out particle and cell sequence is detached and counted, the device can make to be detected when counting
Tally function need not can be realized into detection door and under conditions of realizing that cell to be separated will not be destroyed for cell,
At the detection door of first electric field separates counting channel different sample output passages are accurately flowed under dielectrophoretic force effect.Since separation counts
Channel uses nano-micrometre dimension combination, can greatly reduce required voltage magnitude.
To achieve the goals above, the technical solution of the utility model：
A kind of micro-fluidic chip dress particle being carried out based on nano-micrometre combination of channels and cell sequence is detached and counted
It sets, which includes PDMS micro-fluidic chips, DC power supply, signal amplification circuit and Signal acquiring and processing unit, described
PDMS micro-fluidic chips are to be carved with the side of microchannel in PDMS substrate fovea superiors and glass negative is packaged as a whole, and are waited for forming confession
The micro-fluidic chip of the microchannel of sample circulation, which is characterized in that the microchannel includes：
Sample intake passage, with sample introduction liquid storage hole, the other end is connected with one end of main channel；
Main channel, the other end are connected with sample output passage；
The sample output passage that multiple one end are connected with the main channel respectively, the other end have sample liquid storage hole；
There is first electric field separates counting channel separation to count positive liquid storage hole, and the other end is perpendicular to the main channel
Channel side wall middle section and by first detection door be connected with the main channel；
There is second electric field separates counting channel separation to count cathode liquid storage hole, and the other end is perpendicular to the main channel
The other side channel side wall middle section and by second detection door be connected with the main channel；
And flow field focus channel, with flow field focus channel liquid storage hole, the other end is connected with one end of main channel；
Meanwhile the separation counts positive liquid storage hole, separation counts in cathode liquid storage hole and is inserted with platinum electrode, the direct current
Positive pole is connected with the first electric field separates counting channel, and cathode is connected with the second electric field separates counting channel；Institute
It states DC power supply also to connect a reference resistance, the both ends of the reference resistance connect with the input terminal of the signal amplification component
It connects, the output end of the signal amplification component is connect with signal acquisition control system.
Further, as the preferred of the present invention：
The width of the first detection door is less than the width of the second detection door.
Further, as the preferred of the present invention：
The order of magnitude corresponding to the width of the first detection door is nanoscale, corresponding to the width of the second detection door
The order of magnitude be micron order.
Further, as the preferred of the present invention：
The quantity of the sample output passage is 2.
Compared with prior art, the beneficial effects of the utility model：
1) when being detached, cell to be separated will not be destroyed the utility model when detecting, and can be in dielectrophoresis
Different sample output passages is accurately flowed under force effect；
2) for the utility model when counting, counting need not can be realized into detection door in cell to be separated when detecting
Function；
3) the utility model combines channel by increasing electric field strength ladder when being detached by being provided with nano-micrometre
Degree square enhances dielectrophoretic force, and to effectively prevent, voltage is excessive to cause noise signal that signal-to-noise ratio is caused very much to reduce greatly
Problem；
4) the utility model is easy to operate, and used nanochannel not only may be implemented to detach but also can effectively be counted,
To effectively prevent since two steps are discontinuous or misoperation influences testing result.
Description of the drawings
Fig. 1 is the corresponding structural representation illustration of micro-fluidic chip described in the utility model；
Fig. 2 is the corresponding structural representation illustration of micro flow control chip device described in the utility model.
In figure：M, micro-fluidic chip, L, glass negative, A, sample introduction liquid storage hole, B, flow field focus channel liquid storage hole, C, separation
Counting positive hole, D, separation count cathode hole, E, the first sample outlet hole, F, the second sample outlet hole, 1, main channel, 2, sample intake passage, 3,
Flow field focus channel, 4, separation count positive channel, 5, separation count cathode channel, the 6, first sample output passage, 7, second goes out sample
Channel, the 8, first detection door, the 9, second detection door.
Specific implementation mode
To keep the purpose of this utility model, technical solution and advantage clearer, implement below in conjunction with the utility model
Attached drawing in example, is clearly and completely described the technical solution of the utility model, it is clear that described embodiment is this
Utility model a part of the embodiment, instead of all the embodiments.Based on the embodiments of the present invention, the common skill in this field
The every other embodiment that art personnel are obtained without making creative work belongs to the utility model protection
Range.
Technical background：It can be acted on by dielectrophoretic force when flowing through the place there are electric field due to particle, dielectric constant
Size often determine the Impact direction of particle, and then it is made to flow to different directions；Simultaneously when flowing through the micropore there are electric field
Meeting generates apparent disturbance to electric field so that corresponding micropore both end voltage changes and generates voltage pulse signal, therefore detects
The number of voltage pulse signal, you can realize the number of record sample.
Based on above-mentioned design background, example as shown in Figure 1, the utility model devises a kind of based on nano-micrometre channel
The micro flow control chip device that combination carries out particle and cell sequence is detached and counted includes mainly glass negative L, PDMS miniflow
Control chip M, signal amplification component and the computer as signal acquisition control system；The PDMS micro-fluidic chips be
PDMS substrate fovea superiors are carved with the side of microchannel and glass negative is packaged as a whole, to form the microchannel circulated for sample to be tested
Micro-fluidic chip, which is characterized in that the microchannel includes：
Sample intake passage, with sample introduction liquid storage hole, the other end is connected with one end of main channel；Main channel, the other end
It is connected with sample output passage；The sample output passage that two one end are connected with the main channel respectively i.e. the first sample output passage and second
Sample output passage, the other end have sample liquid storage hole；It hangs down respectively with main channel from the channel side wall middle section of the main channel
Straight electric field separates counting channel；It includes the first electric field separates counting channel and the second electric field separates counting channel, and first
There is electric field separates counting channel separation to count positive liquid storage hole, channel side wall middle part of the other end perpendicular to the main channel
Divide and is connected with the main channel by the first detection door；There is second electric field separates counting channel separation to count cathode liquid storage
Hole, the other end perpendicular to the other side of the main channel channel side wall middle section and by the second detection door and master it is logical
Road is connected；And flow field focus channel, with flow field focus channel liquid storage hole, the other end is connected with one end of main channel
It is logical；Meanwhile the separation counts positive liquid storage hole, separation counts in cathode liquid storage hole and is inserted with platinum electrode, the DC power supply
Positive to be connected with the first electric field separates counting channel, cathode is connected with the second electric field separates counting channel；It is described straight
Galvanic electricity source is also connected a reference resistance, and the both ends of the reference resistance are connect with the input terminal of the signal amplification component,
The output end of the signal amplification component is connect with signal acquisition control system.The work of the corresponding micro flow control chip device
Process：The cell mixing to be separated enters main channel from sample intake passage, then door is detected by described the first/bis-, in dielectric
Swim power under the action of realize separation, simultaneously because cell by it is described the first/bis- detect Men Shihui cause loop voltage change into
And cause the voltage change of reference resistance, it is counted to realize.
Further, as the preferred of the utility model：
The first electric field separates counting channel, the second electric field separates counting channel channel width be 0.75 μm.
Further, as the preferred of the utility model：
The width of the first detection door is less than the width of the second detection door, it is preferred that the width of the first detection door
The corresponding order of magnitude is nanoscale, and the order of magnitude corresponding to the width of the second detection door is micron order.As setting
Nano-micrometre combines channel design, and electric field is largely made to be pooled to a little i.e. so that electric field is applied to nano-micrometre channel
In combination, so that separation and counting is carried out at the same time, do not interfered by pilot process, while the ladder of electric field strength square can be improved
Degree, that is, dielectrophoretic force is enhanced, voltage needed for separation is greatly reduced to ensure that it is (described that cell will not block detection door
The first/bis- detection door does not need particle into that can realize counting, avoids caused by particle enters detection door and blocks), into
And it improves counting precision and optimizes separation and count effect.In addition, having dropwise addition PBS in the sample intake passage and main channel intersection
It flows the side that the flow field focus channel of culture solution detects door to ensure particle on main channel close to above-mentioned the first/bis-.
The preferred signal method element uses difference amplifier element.
The another object of the utility model be to provide for it is a kind of based on the micro flow control chip device carry out cell separation and
The method of counting, which is characterized in that include the following steps：
1) sample is added dropwise：A small amount of PBS buffer solution, which is added dropwise, in each hole of the micro-fluidic chip first makes channel moisten
It is wet, a certain amount of mixing sample is added dropwise in the sample introduction liquid storage hole, is added dropwise in the liquid storage hole of the flow field focus channel appropriate
PBS buffer solution；
2) sample transport：Connect the DC power supply so that the mixing sample in the sample intake passage liquid storage hole is in pressure
It is transported to the main channel under effect, and main channel is pasted by detecting door one under the action of PBS in the focus channel liquid storage hole of flow field
Side wall moves, and particle, due to being acted on by electric field force and pressure, continues when by the first/bis- detection door of vertical main channel
Sample is mixed along straight-ahead motion does not enter flow field focus channel.Sample is mixed in the first/bis- detection door by not Tongfang
To dielectrophoretic force, flow to different sample output passages；
3) signal amplifies collection analysis：The pulse of reference resistance both end voltage is acquired by the platinum electrode of counting channel both sides
Signal is remembered after collected signal is amplified by reference to resistance connection signal amplifier element by signal acquisition control system
It records and shows corresponding detection data, that is, detect the sum of sample；The number detected value of above-mentioned cell is equal to counting channel pulse
The number of signal is equal.
It is detached and is counted by taking polystyrene sample particle and fluorescence granules of polystyrene as an example below, such as Fig. 2 institutes
Show：
The device parameter of the implementation case：Chip sample intake passage used in the implementation case 40*8 μm of size (it is wide ×
It is high), the size of main channel is 120*8 μm (wide × high)；Flow field focus channel size is 120*8 μm (wide × high)；Sample output passage
Size is 120*8 μm (wide × high)；It is 0.75*8 μm (wide × high) to detect door size；Sample to be operated is 5.2 μm of polystyrene
Particle and 7 μm of fluorescence granules of polystyrene mixed solutions；Buffer solution is PBS (1 ×) solution；Be applied to static focusing anode hole and
The voltage in static focusing cathode hole is 80V.
The device includes the sample introduction liquid storage hole A being located on the PDMS micro-fluidic chips, main channel flow field focal aperture B, divides
From count anode hole C, separation counting cathode hole D, the first sample outlet hole E, the second sample outlet hole F and main channel 1, sample intake passage 2,
Flow field focus channel 3, separation count positive channel 4, separation counts cathode channel 5, the first sample output passage 6, the second sample output passage
7, the first detection door 8, the second detection door 9；Wherein it is inserted into respectively between separation counts anode hole C and separation counts cathode hole D
Two platinum electrodes provide 80V voltages；And in one reference resistance R connection of the series connection all the way of 80V voltages；At the both ends reference resistance R
By two conductor in parallel in two input terminals of a difference amplifier；The output end of difference amplifier is connected to data collecting card
And pass through Computer display.
Based on the method that above-mentioned apparatus carries out separation counting, include the following steps：
1) sample is added dropwise：A small amount of PBS buffer solution, which is added dropwise, in each hole of the micro-fluidic chip first makes channel soak,
A certain amount of mixing sample is added dropwise in the sample introduction liquid storage hole A, is added dropwise in the flow field focus channel liquid storage hole B suitable
PBS buffer solution；
2) sample transport：Connect the DC power supply so that the mixing sample in the sample introduction liquid storage hole A is acted in pressure
Under be transported to the main channel 1, and paste main channel by the first detection door 8 under the action of PBS in the focus channel liquid storage hole B of flow field
Either the second detection 9 one side wall of door moves particle when detecting door 9 by the first detection door 8 or second of vertical main channel,
Due to being acted on by electric field force and pressure, continue not entering flow field focus channel 3 along the i.e. mixing sample of straight-ahead motion；Mixing
Sample first detection door 8 either second detection door 9 by the dielectrophoretic force of different directions flow to the first sample output passage 6 or
Second sample output passage 7；The principle of the step is：Since nano-micrometre combination channel has extremely large size gradient, so the
The electric field line that one detection door 8 or second detects door 9 is very intensive, and particle is under the action of flow field focuses PBS buffer solution in mouth
The dielectrophoretic force maximum that particle certifiable in this way is subject to is moved by static focusing mouth one side wall along main channel 1, separating effect reaches
To best；When sample is by detection door, disturbance can be generated to the electric field line for counting mouth both ends, to make voltage change,
The voltage change at the both ends resistance R is further resulted in, detection signal is generated, detection signal is input to acquisition after signal amplifier
Block and shows on computers；
3) signal amplifies collection analysis：The pulse of reference resistance both end voltage is acquired by the platinum electrode of counting channel both sides
Signal is remembered after collected signal is amplified by reference to resistance connection signal amplifier element by signal acquisition control system
It records and shows corresponding detection data, that is, detect the sum of sample；The number detected value of above-mentioned cell is equal to counting channel pulse
The number of signal is equal.Its testing result：Real-time testing result can be directly obtained by computer, and then it is mixed to analyze particle
Close the concrete content of each ingredient in liquid.
The utility model ensures that particle exists in separation process using nano-micrometre combination of channels as shown in the above
The dielectrophoretic force being subject to when separation reaches maximum, reduces required voltage, cell is prevented to be destroyed；In counting process, cell is not required to
To enter detection door, cause count results inaccurate to prevent cell from will detect door blocking, noise signal reduces, and improves letter
It makes an uproar ratio；Simultaneously because two processes are continuously got up, interference and error caused by misoperation are avoided, to effectively raise
Accuracy of detection.
The preferable specific implementation mode of the above, only the utility model, but the scope of protection of the utility model is not
Be confined to this, any one skilled in the art within the technical scope disclosed by the utility model, according to this practicality
Novel technical solution and its inventive concept is subject to equivalent substitution or change, should all cover the scope of protection of the utility model it
It is interior.
Claims (4)
1. a kind of micro flow control chip device particle being carried out based on nano-micrometre combination of channels and cell sequence is detached and counted,
The device includes PDMS micro-fluidic chips, DC power supply, signal amplification circuit and Signal acquiring and processing unit, the PDMS
Micro-fluidic chip is to be carved with the side of microchannel in PDMS substrate fovea superiors and glass negative is packaged as a whole, and test sample is waited for form confession
The micro-fluidic chip of the microchannel of product circulation, which is characterized in that the microchannel includes：
Sample intake passage, with sample introduction liquid storage hole, the other end is connected with one end of main channel；
Main channel, the other end are connected with sample output passage；
The sample output passage that multiple one end are connected with the main channel respectively, the other end have sample liquid storage hole；
There is first electric field separates counting channel separation to count positive liquid storage hole, and the other end is perpendicular to the logical of the main channel
Road side-wall middle section is simultaneously connected by the first detection door with the main channel；
Second electric field separates counting channel, there is separation to count cathode liquid storage hole, and the other end is another perpendicular to the main channel
The channel side wall middle section of side is simultaneously connected by the second detection door with the main channel；
And flow field focus channel, with flow field focus channel liquid storage hole, the other end is connected with one end of main channel；
Meanwhile the separation counts positive liquid storage hole, separation counts in cathode liquid storage hole and is inserted with platinum electrode, the DC power supply
Positive to be connected with the first electric field separates counting channel, cathode is connected with the second electric field separates counting channel；It is described straight
Galvanic electricity source is also connected a reference resistance, and the both ends of the reference resistance are connect with the input terminal of the signal amplification component,
The output end of the signal amplification component is connect with signal acquisition control system.
2. micro flow control chip device according to claim 1, it is characterised in that：
The width of the first detection door is less than the width of the second detection door.
3. micro flow control chip device according to claim 2, it is characterised in that：
The order of magnitude corresponding to the width of the first detection door is nanoscale, the number corresponding to the width of the second detection door
Magnitude is micron order.
4. micro flow control chip device according to claim 1, it is characterised in that：
The quantity of the sample output passage is 2.
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| 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 |
| CN109304094A (en) * | 2018-11-29 | 2019-02-05 | 昆明理工大学 | A kind of active electric diacolation takes the integrating device and its extraction integrated approach of microring array |
| CN109612890A (en) * | 2018-12-28 | 2019-04-12 | 瑞芯智造(深圳)科技有限公司 | A kind of particle counting system and application, the method for detecting metal ion and particle |
| CN111235021A (en) * | 2020-03-06 | 2020-06-05 | 大连海事大学 | Double-liquid-phase separation and detection device and method for circulating tumor cells in peripheral blood |
| CN111644212A (en) * | 2020-05-22 | 2020-09-11 | 华东理工大学 | Micro-fluidic chip and nano-particle separation device |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN109304094A (en) * | 2018-11-29 | 2019-02-05 | 昆明理工大学 | A kind of active electric diacolation takes the integrating device and its extraction integrated approach of microring array |
| CN109612890A (en) * | 2018-12-28 | 2019-04-12 | 瑞芯智造(深圳)科技有限公司 | A kind of particle counting system and application, the method for detecting metal ion and particle |
| CN109612890B (en) * | 2018-12-28 | 2021-02-02 | 瑞芯智造(深圳)科技有限公司 | Particle counting system, application of particle counting system and method for detecting metal ions and particles |
| CN111235021A (en) * | 2020-03-06 | 2020-06-05 | 大连海事大学 | Double-liquid-phase separation and detection device and method for circulating tumor cells in peripheral blood |
| CN111235021B (en) * | 2020-03-06 | 2022-09-27 | 大连海事大学 | Double-liquid-phase separation and detection device and method for circulating tumor cells in peripheral blood |
| CN111644212A (en) * | 2020-05-22 | 2020-09-11 | 华东理工大学 | Micro-fluidic chip and nano-particle separation device |
| CN111644212B (en) * | 2020-05-22 | 2022-05-24 | 华东理工大学 | Micro-fluidic chip and nano-particle separation device |
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