CN111330656A - Micro-fluidic device for micro-particle suspension volume concentration - Google Patents
Micro-fluidic device for micro-particle suspension volume concentration Download PDFInfo
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- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502761—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules
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- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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- 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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- 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
- B01L3/50273—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 means or forces applied to move the fluids
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- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/06—Fluid handling related problems
- B01L2200/0636—Focussing flows, e.g. to laminate flows
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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
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/06—Fluid handling related problems
- B01L2200/0647—Handling flowable solids, e.g. microscopic beads, cells, particles
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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
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- B01L2300/0681—Filter
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
- B01L2300/0883—Serpentine channels
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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
- B01L2400/00—Moving or stopping fluids
- B01L2400/04—Moving fluids with specific forces or mechanical means
- B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
- B01L2400/0409—Moving fluids with specific forces or mechanical means specific forces centrifugal forces
Abstract
The invention discloses a micro-fluidic device for micron particle suspension volume concentration, which consists of a sample inlet, a series of spiral flow channels, a cross-flow filtering flow channel, a Y-shaped branched flow channel, a concentrated sample outlet and a confluence flow channel connected with a blank liquid outlet, wherein the sample inlet is connected with the series of spiral flow channels, the cross-flow filtering flow channel is arranged between the series of spiral flow channels, the tail end of the series of spiral flow channels comprises a sample outlet end and a liquid outlet end, the Y-shaped branched flow channel is connected with the sample outlet end at the tail end of the series of spiral flow channels, the concentrated sample outlet is connected with the Y-shaped branched flow channel, and the confluence flow channel is connected with the liquid outlet. The device can realize high flux and high rate concentration of micron particle suspension, the optimal recovery rate is up to 99.99%, accurate flow resistance matching is not needed, the operation is simple, meanwhile, the damage to cells is low, the activity of the treated cells is up to 95%, the structure is simple, and the cost is low.
Description
Technical Field
The present invention relates to a volume-concentrating microfluidic device, and more particularly, to a microparticle suspension volume-concentrating microfluidic device.
Background
Biological suspensions containing cells or particles are important medical samples, counting and detecting of the cells or particles in the biological suspensions can provide important information for disease diagnosis, concentration pretreatment needs to be carried out on the samples to realize detection of the cells or particles in the biological suspensions, and detection sensitivity and detection rate are improved by means of concentration increase of target components in the samples, and a currently common sample concentration method comprises the following steps: high speed centrifugation and membrane filtration. Centrifugation forces cells or particles to settle to the bottom of a centrifuge tube by strong centrifugal force, followed by reduction of the volume of the sample by removal of the supernatant, resulting in an increase in cell or particle concentration. However, the centrifugal method is expensive in equipment, cannot be used in complex working conditions such as the field and the like, cannot be used for efficiently concentrating samples with extremely low cell/particle concentration or samples with ultra-large volume, and can cause damage and death of biological particles such as cells and the like due to strong centrifugal force. The method is simple and direct, but the cell blocked on the filter membrane is difficult to recover, the blockage problem of the filter membrane can cause that the efficiency is rapidly reduced along with the use time, and the fixed filter pores are only suitable for filtering and selecting the cells or the particles with specific sizes, so that the application range is small.
Disclosure of Invention
The purpose of the invention is as follows: the invention aims to provide a micro-fluidic device for micro-particle suspension volume concentration, which can realize high-throughput and high-rate concentration, has low cell damage, simple operation, strong universality and low cost.
The technical scheme is as follows: the micro-fluidic device for micron particle suspension volume concentration comprises a sample inlet, a series of spiral flow channels, a cross flow filtering flow channel, a Y-shaped branched flow channel, a concentrated sample outlet and a confluence flow channel connected with a blank liquid outlet, wherein the sample inlet is connected with the series of spiral flow channels, the cross flow filtering flow channel is arranged between the series of spiral flow channels, the tail end of the series of spiral flow channels comprises a sample outlet end and a liquid outlet end, the Y-shaped branched flow channel is connected with the sample outlet end at the tail end of the series of spiral flow channels, the concentrated sample outlet is connected with the Y-shaped branched flow channel, and the confluence flow channel is connected with the liquid outlet end at the.
The cross section of the series of spiral flow channels is a rectangular cross section, the height h of the cross section is less than or equal to 100d/7, a flow resistance flow channel is connected between the outlet of the concentrated sample and the Y-shaped branched flow channel, the flow resistance flow channel is an S-shaped flow resistance flow channel unit or a sine line-shaped concentrated flow channel, the tail end of the sine line-shaped concentrated flow channel is connected with a cross-shaped three-branched outlet system, the series of spiral flow channels comprise an outermost spiral flow channel, a second spiral flow channel and an innermost spiral flow channel which are parallel to each other, and the outermost spiralThe circle spiral flow channels are connected through a cross flow filtering flow channel, the second circle spiral flow channel is connected with the innermost circle spiral flow channel through a cross flow filtering flow channel, the outermost circle spiral flow channel is connected with a sample inlet, the outermost circle spiral flow channel is connected with the tail end of the second circle spiral flow channel to form a liquid outlet end, and the tail end of the innermost circle spiral flow channel is a sample outlet end; the second circle of spiral flow channel starts from the spiral winding position of the outermost circle of spiral flow channel by 0.5-1.5 circles, and the innermost circle of spiral flow channel starts from the spiral winding position of the second circle of spiral flow channel by 0.5-1 circles; the included angles between the cross-flow filtering flow channel and the outermost circle of spiral flow channel, between the second circle of spiral flow channel and the innermost circle of spiral flow channel are acute angles; the rectangular section width relation of the outermost ring spiral flow channel, the second ring spiral flow channel and the innermost ring spiral flow channel is W2≧W3≧W4。
Has the advantages that: compared with the prior art, the invention has the following remarkable advantages: 1. the high-flux and high-rate concentration of the micron particle suspension can be realized, and the optimal recovery rate is up to 99.99%; 2. accurate flow resistance matching is not needed, and the operation is simple; 3. the damage to cells is low, and the activity of the treated cells is as high as 95 percent; 4. simple structure and low cost.
Drawings
FIG. 1 is a schematic structural view of example 1;
FIG. 2 is a schematic diagram of the migration movement of microparticles in the outermost spiral flow channel to the second spiral flow channel;
FIG. 3 is a schematic diagram of the migration motion of microparticles in the second spiral flow path to the innermost spiral flow path;
FIG. 4 is a schematic structural view of example 3;
FIG. 5 is the results of the concentration performance test of example 1;
FIG. 6 is the results of the concentration performance test of example 2;
FIG. 7 shows the results of the concentration performance of example 1 on leukocyte suspensions and example 2 on MCF-7 cell suspensions;
FIG. 8 is a comparison of the leukocyte suspensions of example 1 before and after concentration;
FIG. 9 is a graph showing the results of example 1 in which the leukocyte suspension was concentrated 3 times.
Detailed Description
Example 1
As shown in fig. 1, the micro-fluidic device for micro-particle suspension volume concentration is composed of a sample inlet 1, a series of spiral flow channels, a cross-flow filtration flow channel 5, a Y-shaped branched flow channel 6, a concentrated sample outlet 8 and a confluence flow channel 9 connected with a blank liquid outlet 10, wherein the sample inlet 1 is connected with the series of spiral flow channels, the cross-flow filtration flow channel 5 is arranged between the series of spiral flow channels, the tail end of the series of spiral flow channels comprises a sample outlet end and a liquid outlet end, the Y-shaped branched flow channel 6 is connected with the sample outlet end at the tail end of the series of spiral flow channels, the concentrated sample outlet 8 is connected with the Y-shaped branched flow channel 6 through an S-shaped flow resistance flow channel unit 7, and; the section of the series spiral flow channel is a rectangular section, the height of the section is 100 mu m, the series spiral flow channel is composed of an outermost circle of spiral flow channel 2, a second circle of spiral flow channel 3 and an innermost circle of spiral flow channel 4 which are parallel to each other, the outermost circle of spiral flow channel 2 and the second circle of spiral flow channel 3 are connected through a cross flow filtering flow channel 5, the second circle of spiral flow channel 3 and the innermost circle of spiral flow channel 4 are connected through the cross flow filtering flow channel 5, the outermost circle of spiral flow channel 2 is connected with a sample inlet 1, the outermost circle of spiral flow channel 2 and the tail end of the second circle of spiral flow channel 3 are connected together to form a liquid outlet end, the tail end of the innermost circle of spiral flow channel 4 is a sample outlet end, the second circle of spiral flow channel 3 starts from the outermost circle of spiral flow channel 2 to be spirally wound for 1 circle, the innermost circle of spiral flow channel 4 starts from the second circle of spiral flow channel 3 to be spirally, The included angle between the second circle of spiral flow passage 3 and the innermost circle of spiral flow passage 4 is 60 degrees, the rectangular cross section widths of the outer circle of spiral flow passage 2, the second circle of spiral flow passage 3 and the innermost circle of spiral flow passage 4 are W respectively2＝600μm，W3＝500μm，W4＝400μm。
In the test, sample liquid to be concentrated of white blood cell WBC with the particle size of 10 μm or 15 μm and the diameter of 10 μm is prepared, the sample liquid to be concentrated of the particle size of 10 μm or 15 μm is made of polystyrene particles, a micro particle suspension volume concentration micro-fluidic device is injected from a sample inlet 1 by virtue of a syringe pump at the volume flow rates of 3ml/min, 3.5ml/min, 4ml/min, 4.5ml/min, 5ml/min, 5.5ml/min and 6ml/min, as shown in FIG. 2, the spiral flow channel 2 at the outermost circle is communicated with the spiral flow channel 3 at the second circle by virtue of a cross-flow filtering flow channel 5, particle beams formed by focusing are gradually transferred from the spiral flow channel 2 at the outermost circle to the spiral flow channel 3 at the second circle by virtue of the cross-flow filtering flow channel 5, the micro particles transferred to the spiral flow channel 3 at the second circle are transferred along the spiral flow channel 3, and is refocused to form a column in the second circle of spiral flow channel 3, as can be seen from fig. 3, the micron particles focused into a bundle are transferred to the innermost circle of spiral flow channel 4 by the series of cross-flow filtration flow channels 5 under the cross-flow filtration effect, so that the micron particles are all transferred to the innermost circle of spiral flow channel 4 from the outermost circle of spiral flow channel 2, and only blank liquid without particles remains in the outermost circle of spiral flow channel 2 and the second circle of spiral flow channel 3, thereby realizing the separation of the micron particles and the blank liquid, concentrating the suspended micron particles in the sample, refocusing the micron particles to be close to the side wall surface of the center of the spiral to form a bundle in the innermost circle of spiral flow channel 4, further removing the liquid close to the outer side in the flow channel by the Y-shaped branched flow channel 6, reducing the volume of the concentrated sample by the S-shaped blank flow resistance flow channel unit 7, improving the concentration effect, and leading out the concentrated micron particle sample from the, and the blank liquid in the outermost spiral flow passage 2 and the second spiral flow passage 3 and the blank liquid generated by the branch outside the Y-shaped branched flow passage 6 are collected in the confluence flow passage 9 and are led out and collected by a blank liquid outlet 10.
As shown in FIG. 5(a), the optimum concentration effect was obtained at a volume flow rate of 4ml/min for the 10 μm particles, the optimum concentration ratio was 12.21, the optimum concentration ratio was 12.31 for the 15 μm particles at a volume flow rate of 4.5ml/min, the optimum recovery rate was 97.67% for the 10 μm particles and 98.45% for the 15 μm particles, as shown in FIG. 5 (b).
Using a micro-particle suspension volume concentration micro-fluidic device to perform three times of concentration on 490 milliliters of leukocyte biological suspension at a volume flow rate of 4ml/min, wherein microscope sampling pictures before and after concentration are shown in fig. 8(a) and (b), the leukocyte concentration in a sample after concentration is greatly improved, as shown in fig. 9, I represents an initial sample, R1-R3 respectively represent three times of concentration, and the leukocyte concentration in a target sample is 1.17 × 104The volume/ml is lifted to 1.27 × 107Is/areMl, and the integral concentration ratio is up to 1085 times.
Example 2
This example is different from example 1 in that the height of the cross section of the serial spiral flow channel is 150 μm, the sample solution to be concentrated of 15 μm and 20 μm particles and the human breast cancer cells MCF-7 with the diameter of 20 μm is prepared, and the microparticle suspension volume concentration microfluidic device is injected from the sample inlet 1 by means of a syringe pump at the volume flow rates of 3ml/min, 3.5ml/min, 4ml/min, 4.5ml/min, 5ml/min, 5.5ml/min and 6 ml/min.
As shown in FIG. 6(a), the 15 μm particles can achieve the best concentration effect at a volume flow rate of 4ml/min, the best concentration ratio is 12.17, the best concentration ratio of 20 μm particles at a volume flow rate of 5ml/min is 12.49, the best recovery rate from FIG. 6(b) is 97.33% for 15 μm particles, and the best recovery rate from 20 μm particles is 99.99%.
As shown in FIG. 7(a), the optimal concentration ratio of example 1 to leukocytes at a volume flow rate of 4ml/min was 10.79, and the optimal concentration ratio of example 2 to MCF-7 cells at a volume flow rate of 4ml/min was 11.30, as shown in FIG. 7(b), the optimal recovery rate of example 1 to leukocytes was 87.97%, the optimal recovery rate of example 2 to MCF-7 cells was 90.40%, and the activity of the cells after the device treatment was still greater than 95%, indicating low cell damage and high biocompatibility of the device.
Example 3
The difference between this embodiment and embodiment 1 is that the concentrated sample outlet 8 and the Y-shaped branched flow channel 6 are connected through a sinusoidal line type concentrated flow channel 13 and a cross-shaped three-branched outlet system 14, the sinusoidal line type concentrated flow channel 13 is used to focus the micrometer particles to the middle area of the flow channel, the three-branched outlet system 14 is used to remove the blank liquid on both sides, and the concentration effect is further improved, and the structure is shown in fig. 4.
The device is used for concentrating concentrated sample liquid of MCF-7 cells and A549 lung cancer cells, the concentration of the MCF-7 cells obtains the best concentration effect under the volume flow of 4ml/min, the best concentration ratio is 41, the best concentration ratio of the A549 lung cancer cells under the volume flow of 4ml/min is 41.2, the best recovery rate of the MCF-7 cells is 89.5%, and the best recovery rate of the A549 lung cancer cell particles is 90%.
Claims (10)
1. The micro-fluidic device is characterized by comprising a sample inlet (1), a series of spiral flow channels, a cross-flow filtering flow channel (5), a Y-shaped branched flow channel (6), a concentrated sample outlet (8) and a confluence flow channel (9) connected with a blank liquid outlet (10), wherein the sample inlet (1) is connected with the series of spiral flow channels, the cross-flow filtering flow channel (5) is arranged between the series of spiral flow channels, the tail end of the series of spiral flow channels comprises a sample outlet end and a liquid outlet end, the Y-shaped branched flow channel (6) is connected with the sample outlet end at the tail end of the series of spiral flow channels, the concentrated sample outlet (8) is connected with the Y-shaped branched flow channel (6), and the confluence flow channel (9) is connected with the liquid outlet end at the tail end of the series of spiral flow channels.
2. The microparticle suspension volume concentration microfluidic device according to claim 1, wherein the series of spiral flow channels have a rectangular cross section, a cross section height h ≦ 100d/7, and d is microparticle diameter.
3. The microparticle suspension volume concentration microfluidic device according to claim 1, wherein a flow resistance channel is connected between the concentrated sample outlet (8) and the Y-shaped branched channel (6).
4. A microparticle suspension volume concentration microfluidic device according to claim 3, wherein said flow resistance channel is an S-shaped flow resistance channel unit (7).
5. The microparticle suspension volume concentration microfluidic device according to claim 3, wherein the flow resistance channel is a sinusoidal line type concentration channel (13).
6. The micro fluidic device for micro particle suspension volume concentration according to claim 5, wherein the sinusoidal line-shaped concentration flow channel (13) is connected with a cross-shaped three-split outlet system (14) at the end.
7. The microparticle suspension volume concentration microfluidic device according to claim 1, wherein the series of spiral flow channels comprises an outermost spiral flow channel (2), a second spiral flow channel (3) and an innermost spiral flow channel (4) which are parallel to each other, the outermost spiral flow channel (2) and the second spiral flow channel (3) are connected through a cross-flow filtration flow channel (5), the second spiral flow channel (3) and the innermost spiral flow channel (4) are connected through a cross-flow filtration flow channel (5), the outermost spiral flow channel (2) is connected with the sample inlet (1), the outermost spiral flow channel (2) and the second spiral flow channel (3) are connected together at their ends to form a liquid outlet, and the innermost spiral flow channel (4) is connected with the sample outlet.
8. The microparticle suspension volume concentration microfluidic device according to claim 4, wherein the second spiral channel (3) starts from the outermost spiral channel (2) and spirals 0.5-1.5 times, and the innermost spiral channel (4) starts from the second spiral channel (3) and spirals 0.5-1 times.
9. The micro-fluidic device for micro-particle suspension volume concentration according to claim 4, wherein the cross-flow filtration flow channel (5) forms an acute angle with the outermost spiral flow channel (2), the second spiral flow channel (3) and the innermost spiral flow channel (4).
10. The microparticle suspension volume concentration microfluidic device according to claim 4, wherein the rectangular cross-sectional width relationship of the outermost spiral flow channel (2), the second spiral flow channel (3) and the innermost spiral flow channel (4) is W2≧W3≧W4。
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| CN112452364A (en) * | 2020-11-18 | 2021-03-09 | 江南大学 | Micro-fluidic chip for rapid sorting and manufacturing method |
| CN112547145A (en) * | 2020-11-19 | 2021-03-26 | 东南大学 | Rare cell rapid screening micro-fluidic device |
| CN113522383A (en) * | 2021-06-25 | 2021-10-22 | 东南大学 | Cell working condition device |
| WO2022161371A1 (en) * | 2021-01-29 | 2022-08-04 | 广州万孚生物技术股份有限公司 | In vitro analysis diagnostic instrument, circulating tumor cell sorting and enrichment micro-fluidic chip and method |
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