CN111040938A - Microfluidic chip and sorting method - Google Patents

Microfluidic chip and sorting method Download PDF

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CN111040938A
CN111040938A CN201911240815.6A CN201911240815A CN111040938A CN 111040938 A CN111040938 A CN 111040938A CN 201911240815 A CN201911240815 A CN 201911240815A CN 111040938 A CN111040938 A CN 111040938A
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sorting
channel
flow channel
inlet
main
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陈华英
陈震林
朱永刚
徐东
赵力涛
徐建华
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Shenzhen Graduate School Harbin Institute of Technology
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    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/02—Form or structure of the vessel
    • C12M23/16—Microfluidic devices; Capillary tubes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
    • C12M33/14—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus with filters, sieves or membranes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
    • C12M47/04—Cell isolation or sorting

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Abstract

The invention discloses a micro-fluidic chip and a sorting method based on the cell sorting micro-fluidic chip. The microfluidic chip comprises a channel layer, wherein the channel layer comprises a main runner for sample liquid to pass through; the separation flow channel is communicated with the main flow channel through a separation flow channel inlet, and the height of the separation flow channel inlet is smaller than that of the main flow channel. When the sample liquid reaches the intersection of the separation flow channel and the main flow channel, the particles with the particle size not larger than the inlet height of the separation flow channel enter the separation flow channel, and the particles with the larger particle size continuously flow in the main flow channel, so that the particles with different sizes are separated. According to the method, the height of the sorting flow channel can be dynamically changed in the using process so as to change the diameter of particles entering the sorting flow channel, the purpose of screening different-size particles by the same device is achieved, and efficient sorting can be achieved by setting the heights of the main flow channel and the sorting flow channel in advance without manual intervention in the sorting process.

Description

Microfluidic chip and sorting method
Technical Field
The invention relates to the technical field of microfluidic chips, in particular to a microfluidic chip and a sorting method.
Background
In recent years, with the progress of micro-nano manufacturing technology and fluid precise control technology, new microfluidic and lab-on-a-chip technologies have become more and more important as efficient cell and particle separation and sorting methods. The microfluidic technology can greatly reduce the expensive sample size and realize high-resolution separation of various cells until the single cell level. Different types of cells have very different physical properties, such as size, shape and even deformability. On this basis, microfluidic chips have emerged that sort cells according to their different physical characteristics. Peter Wilding achieved red blood cell enrichment using a filter array, but this method resulted in red blood cells becoming blocked in the channel and failing to allow more processing analysis of the cells. Daniel Lee sets the filter array in a slanted arrangement in the microchannel to allow unfiltered residues to travel with the mainstream while the target cells are sorted to another channel and collected, thereby preventing cells from accumulating clogging at the filter. However, although this device solves the clogging problem of the filter array, the filtration efficiency is to be improved. Some microfluidic chips introduce manual intervention to improve the sorting efficiency, such as separation and sorting of cells with antibody-labeled magnetic beads, but these methods may interfere with the sorting result to some extent.
Disclosure of Invention
The present invention is directed to solving at least one of the problems of the prior art. Therefore, the invention provides a micro-fluidic chip which can realize high-efficiency sorting without manual intervention on sample liquid.
The invention also provides a sorting method based on the sorting microfluidic chip.
In a first aspect, an embodiment of the present invention provides a microfluidic chip, including a channel layer, where the channel layer includes: the main flow passage is used for passing the sample liquid; the separation flow channel is communicated with the main flow channel through a separation flow channel inlet, and the height of the separation flow channel inlet is smaller than that of the main flow channel.
The micro-fluidic chip provided by the embodiment of the invention at least has the following beneficial effects:
when the sample liquid reaches the intersection of the sorting flow channel and the main flow channel, cells or other particles with the particle size not larger than the height of the inlet of the sorting flow channel can enter the sorting flow channel, and the cells or other particles with the particle size larger than the height of the sorting flow channel can continuously flow in the main flow channel along the outer wall of the intersection under the action of the sample liquid, so that the cells or other particles with different sizes are sorted out. According to the micro-fluidic chip, the heights of the main flow channel and the sorting flow channel can be set in advance through a micro-nano technology, and efficient sorting can be realized without manual intervention in the sorting process.
According to the microfluidic chip of further embodiments of the present invention, the main channel is connected to the sorting channel at both sides in the axial direction thereof through the sorting channel inlet. Because the single separation flow channel can only be arranged on one side of the main flow channel, the sample liquid and the particles contained in the sample liquid are in a laminar flow state in the flow channel, and the sample liquid on one side far away from the separation flow channel in the main flow channel can not be separated through the inlet of the separation flow channel. And through set up separation runner entry intercommunication separation runner in the both sides of sprue, can guarantee that the particle along the different distances parallel flow from sprue both sides wall can both be sorted out, and the quantity of separation runner can show the efficiency that improves the separation.
In a second aspect, an embodiment of the present invention provides a microfluidic chip, including:
the channel layer comprises a main flow channel for sample liquid to pass through and a separation flow channel, and the separation flow channel is communicated with the main flow channel through a separation flow channel inlet;
the control layer is stacked on at least one side of the channel layer, a pressure pipeline is arranged on the control layer corresponding to the position of the sorting flow channel inlet and used for driving the sorting flow channel inlet to shrink in the height direction, and therefore the height of the sorting flow channel inlet is smaller than that of the main flow channel.
The micro-fluidic chip provided by the embodiment of the invention at least has the following beneficial effects:
through the setting of pressure conduit in the control layer, can be less than the height of mainstream way within range nimble height of adjusting the sorting runner entry according to the size of the cell of waiting to sort or other microparticle to make it can adapt to different sorting demands.
In addition, cells or other particles of a specific size in the sample solution flow into the sorting channel with a certain probability and are collected separately. On the basis of the sorting flow channel, the total sorting efficiency can be effectively improved by simply increasing the number of sorting flow channel inlets (namely, increasing the number of sorting flow channels).
According to the microfluidic chip of other embodiments of the present invention, the pressure conduit may pneumatically or hydraulically adjust the height of the inlet of the sorting channel.
According to the microfluidic chip of other embodiments of the present invention, a thin film layer is further disposed between the control layer and the channel layer, the thin film layer is provided with an elastic membrane, and the pressure pipeline drives the elastic membrane to deform in a height direction of the inlet of the sorting flow channel so as to adjust the height of the inlet of the sorting flow channel. The pressure pipeline of the control layer controls the deformation degree of the elastic membrane, and when the pressure in the pressure pipeline changes, the elastic membrane of the thin film layer deforms towards the sorting flow channel direction, so that the height of the inlet of the sorting flow channel changes. The height of the inlet of the sorting flow channel can be accurately controlled by controlling the pressure of the pressure pipeline, so that the sorting channel is controlled to sort cells or other particles with different sizes in the sample liquid.
According to other embodiments of the microfluidic chip of the present invention, the pressure channel is inclined to a flow direction of the main channel. After the pressure pipeline is used for pressing the thin film layer, cells or other particles with the particle size larger than the height of the inlet of the sorting flow channel in the sample liquid can be ensured to be prevented from being blocked at the inlet of the sorting flow channel as far as possible, and the cells or other particles can continuously enter the main flow channel under the driving of the sample liquid.
According to the microfluidic chip of other embodiments of the present invention, the elastic membrane may be disposed only at a position on the thin film layer corresponding to the inlet of the sorting channel or may be disposed on the entire thin film layer, and only the pressure channel is disposed at a position on the control layer corresponding to the inlet of the sorting channel, so that when the pressure of the pressure channel is changed, the height of the inlet of the sorting channel is changed within a range smaller than the height of the main channel, and the specific height of the inlet of the sorting channel is flexibly adjusted by the pressure channel.
According to other embodiments of the microfluidic chip of the present invention, the main channel is connected to the sorting channel through the sorting channel inlet at both sides in the axial direction thereof. Because the single sorting flow channel can only be arranged on one side of the main flow channel, the sample liquid which is far away from one side of the sorting flow channel in the main flow channel can not be sorted through the inlet of the sorting flow channel. And through set up sorting flow channel entry intercommunication sorting flow channel in the both sides of sprue, can further guarantee that the cell or other particle of specific size can both be sorted out by the separation in the sample liquid that gets into the sprue and select separately, improve the efficiency of selecting separately.
According to the microfluidic chip of the other embodiments of the present invention, the sorting channel inlets of both sides of the main channel are staggered.
The sorting flow channel inlets are arranged on two sides of the main flow channel in a staggered mode, so that cells or other particles with specific sizes in the sample liquid in the main flow channel can be timely sorted out.
According to the microfluidic chip of other embodiments of the present invention, the main channel is further provided with a filtering mechanism. The filtering mechanism is arranged on the main runner, and particularly the filtering mechanism is arranged before the inlet of the first sorting runner on the main runner, so that impurities with overlarge volumes can be filtered, and the main runner and the sorting runner are prevented from being blocked by the impurities.
According to the microfluidic chip of other embodiments of the present invention, the filtering mechanism may be a plurality of columns arranged along the flow direction, and a filtering gap capable of blocking impurities with large volume is formed between the columns.
According to the microfluidic chip of the other embodiments of the present invention, the sorting channels are arranged in parallel.
In a third aspect, an embodiment of the present invention provides a sorting method using the above microfluidic chip, including the following steps:
s1: driving the inlet of the sorting flow channel to contract in the height direction;
s2: inputting sample liquid into the main flow channel;
s3: and respectively collecting the sample liquid and the particles contained in the sample liquid at the outlet of the main flow channel and the outlet of the sorting flow channel.
The method for sorting by applying the microfluidic chip provided by the embodiment of the invention at least has the following beneficial effects:
driving the sorting flow inlet on the microfluidic chip to contract in the height direction of the sorting flow inlet, so that the sorting flow inlet is adjusted to allow the entry of cells or other particles with specific sizes; then introducing sample liquid into the main flow channel; cells or other particles with specific sizes can be efficiently sorted out by the sorting flow channel without manual intervention interference on sample liquid, and the high efficiency of sorting is ensured.
Drawings
Fig. 1 is a schematic view of a channel layer of a microfluidic chip according to an embodiment of the present invention;
fig. 2 is an enlarged view of a position a of a channel layer of the microfluidic chip shown in fig. 1;
fig. 3 is a schematic diagram of a control layer of a microfluidic chip according to another embodiment of the present invention;
FIG. 4 is a schematic view of the microfluidic chip shown in FIG. 3;
FIG. 5 is an enlarged view of the microfluidic chip shown in FIG. 4 at position B;
FIG. 6a is a cross-sectional side view of the microfluidic chip shown in FIG. 4 at position B when the pressure line is not pressurized;
FIG. 6B is a cross-sectional side view of the microfluidic chip shown in FIG. 4 at position B under pressure applied by the pressure line;
fig. 7 is a partial schematic view of a microfluidic chip according to yet another embodiment of the present invention.
Detailed Description
The concept and technical effects of the present invention will be clearly and completely described below in conjunction with the embodiments to fully understand the objects, features and effects of the present invention. It is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments, and those skilled in the art can obtain other embodiments without inventive effort based on the embodiments of the present invention, and all embodiments are within the protection scope of the present invention.
In the description of the embodiments of the present invention, if an orientation description is referred to, for example, the orientations or positional relationships indicated by "upper", "lower", "front", "rear", "left", "right", etc. are based on the orientations or positional relationships shown in the drawings, only for convenience of describing the present invention and simplifying the description, but not for indicating or implying that the referred device or element must have a specific orientation, be constructed and operated in a specific orientation, and thus should not be construed as limiting the present invention.
In the description of the embodiments of the present invention, if a feature is referred to as being "disposed", "fixed", "connected", or "mounted" to another feature, it may be directly disposed, fixed, or connected to the other feature or may be indirectly disposed, fixed, connected, or mounted to the other feature. In the description of the embodiments of the present invention, if "a number" is referred to, it means one or more, if "a plurality" is referred to, it means two or more, if "greater than", "less than" or "more than" is referred to, it is understood that the number is not included, and if "greater than", "lower" or "inner" is referred to, it is understood that the number is included. If reference is made to "first" or "second", this should be understood to distinguish between features and not to indicate or imply relative importance or to implicitly indicate the number of indicated features or to implicitly indicate the precedence of the indicated features.
Example 1
Referring to fig. 1 and 2, fig. 1 is a schematic view of a channel layer of a microfluidic chip according to an embodiment of the present invention, and fig. 2 is an enlarged view of a position a of the channel layer shown in fig. 1. The channel layer comprises a sample liquid inlet 110 and a filtering part 120 communicated with the sample liquid inlet 110, wherein the filtering part 120 is a plurality of columnar body layers, each columnar body layer is provided with a plurality of columnar bodies arranged at intervals, and the intervals can intercept impurities with larger volume so as to avoid blocking the impurities after entering the main flow passage 130. A main flow channel 130 is connected to the rear of the filter unit 120, and a sample liquid outlet 150 is provided at the end of the main flow channel 130. A plurality of separation channel inlets 141 are further provided at intervals at both sides of the main channel 130, and are communicated with the separation channel 140 through the separation channel inlets 141, and a separation outlet 160 is provided at the end of the separation channel 140. The height of the sorting channel inlet 141 is different from that of the main channel 130, and specifically, when the channel layer of the microfluidic chip of this embodiment is prepared by using the micro-nano technology, the height of the sorting channel inlet 141 is controlled to be 15 μm, and the height of the main channel 130 is controlled to be 50 μm. When the microfluidic chip is used for cell sorting, a sample liquid is injected into the sample liquid inlet 110, impurities with overlarge volume are intercepted at the filtering part 120, and after the cells to be sorted enter the main flow channel 130, the cells or other particles with the particle size larger than the height of the sorting flow channel inlet 141 (namely the particle size is larger than 15 microns) cannot enter the sorting flow channel 140 and are kept in the main flow channel 130 to move and are finally discharged or further collected from the sample liquid outlet 150 when the cells or other particles contact the sorting flow channel inlet 141; and cells or other fine particles having a particle size not greater than the height of the sorting flow channel inlet 141 (i.e., having a particle size not greater than 15 μm) can enter the sorting flow channel 140 and be collected via the sorting outlet 160 when contacting the sorting flow channel inlet 141 repeatedly disposed at both sides of the main flow channel 130, thereby enabling the cells to be sorted according to their sizes.
Example 2
Referring to fig. 3 to 5, fig. 3 is a schematic view of a control layer of a microfluidic chip according to another embodiment of the present invention, fig. 4 is a schematic view of the microfluidic chip shown in fig. 3, and fig. 5 is an enlarged view of a position B of the microfluidic chip shown in fig. 4. The control layer of the microfluidic chip includes a plurality of pressure conduits 310 arranged in parallel, the ends of the pressure conduits 310 are provided with gas source inlets 320, and pressure of the pressure conduits 310 is adjusted through the gas source inlets 320, that is, in this embodiment, the pressure conduits 310 are air pressure conduits. The channel layer of the microfluidic chip comprises a sample liquid inlet 110 and a filter part 120 communicated with the sample liquid inlet 110. A main flow channel 130 is connected to the rear of the filter unit 120, and a sample liquid outlet 150 is provided at the end of the main flow channel 130. A plurality of separation channel inlets 141 are further provided at intervals at both sides of the main channel 130, and communicate with the separation channel 140 through the separation channel inlets 141. The end of the sort flow channel 140 is provided with a sort outlet 160. After the control layer is combined with the channel layer, the contact end 311 of the pressure conduit 310 is disposed at the location of the sort flow channel inlet 141 of the sort flow channel 140. The arrows in fig. 5 indicate the flow direction of the sample liquid in the main channel 130, and the contact end 311 of the pressure tube 310 is inclined to the flow direction of the sample liquid in the main channel 130, so that when the sample liquid passes through the sorting channel inlet 141, cells with a larger particle size are ensured to flow downstream with the push of the sample liquid rather than being blocked at the sorting channel inlet 141. A thin film layer (not shown) is also provided between the control layer and the channel layer of the microfluidic chip.
Referring to fig. 6a and 6B, fig. 6a is a cross-sectional side view of the microfluidic chip shown in fig. 4 at position B when the pressure channel is not pressurized; fig. 6B is a cross-sectional side view of the microfluidic chip shown in fig. 4 at position B when the pressure channel is pressurized. When the pressure pipe 310 is not pressurized, the initial height of the sorting flow channel inlet 141 is the same as the height of the main flow channel 130; when the pressure pipe 310 presses downward (i.e., the position of the inlet 141 of the separation flow channel), the elastic membrane 200 of the thin film layer deforms downward, and the height of the inlet 141 of the separation flow channel changes, which has a certain height difference from the main flow channel 130. When the sample solution is introduced into the main channel 130 and passes through the position B, the cells having a particle diameter larger than the height of the sorting channel inlet 141 in the sample solution on the left side in the flow direction are pushed by the sample solution along the height change boundary by the sorting channel inlet 141 and flow downstream in the part of the main channel 130 having a higher channel height. Because the cells move along the streamlines in the laminar flow, whether a cell can be sorted at the sorting flow-channel inlet 141 depends primarily on whether the streamlines in which the cell is located flow toward the sorting flow-channel inlet 141. If the streamlines in which the particles are located in the higher level portion of the primary channel 130, the cells continue to flow downstream. An identical and symmetrical sorting channel inlet 141 is therefore provided downstream of the primary channel 130 to ensure that all particles have the opportunity to be sorted at the sorting channel inlet 141. At each such sort channel inlet 141, cells of a particular size in the sample fluid flow into the sort channel 140 with a certain probability and are collected through the sort outlet 160. Simply increasing the number of sorting channel inlets 141 and sorting channels 140 can effectively increase the overall efficiency of sorting.
Example 3
Referring to fig. 7, which is a partial schematic view of a microfluidic chip according to still another embodiment of the present invention, the microfluidic chip is different from embodiment 2 in that a main channel 130 of a channel layer, a sorting channel 140, and a pressure channel 310 of a control layer form three sections including a first sorting region a, a second sorting region b, and a third sorting region c, the sorting channels 140 arranged in parallel on one side in the first sorting region a lead to a first sorting port 160a, and the corresponding pressure channels 310 communicate with a first gas source inlet 320 a; similarly, the parallel sorting channels 140 on one side in the second sorting zone b lead to the second sorting opening 160b, and the corresponding pressure pipes 310 are communicated with the second gas source inlet 320 b; the side-by-side sorting channels 140 in the third sorting zone c lead to the third sorting opening 160c, and the corresponding pressure conduits 310 communicate with the third gas supply inlet 320 c. The first air source inlet 320a, the second air source inlet 320b, and the third air source inlet 320c can respectively introduce different amounts of air to form different-sized extrusion on the elastic membrane, so that the heights of the sorting channel inlets in the first sorting area a, the second sorting area b, and the third sorting area c are in different distribution, specifically, da<db<dc<dMaster and slave(da、db、dc、dMaster and slaveRespectively representing the heights of the sorting channel inlet of the first sorting region a, the sorting channel inlet of the second sorting region b, the sorting channel inlet of the third sorting region c and the main channel, namely the heights of the sorting channel inlet of the first sorting region a, the sorting channel inlet of the second sorting region b, the sorting channel inlet of the third sorting region c and the main channel are sequentially increased, and the sizes of the cells or other particles to be sorted collected through the first sorting outlet 160a, the second sorting outlet 160b, the third sorting outlet 160c and the sample liquid outlet are respectively (0, d)a]、(da,db]，(db,dc]，(dc,dMaster and slave]Wherein "(" means not including the size "]"means including the size.
The embodiment can realize that when the sample liquid flows in, different sorting areas of the microfluidic chip can sort and enrich cells or other particles with different sizes. The height of the inlet of the sorting flow channel can be accurately controlled by controlling the pressure of the pressure pipeline, and cells or other particles with different sizes in the sorting sample liquid of the sorting flow channel are controlled.
Compared with other existing sorting methods and sorting devices, in the above embodiment of the invention, fewer original samples are required during sorting and separation, the sorting efficiency is higher, and artificial interference such as antibodies and magnetic beads is not required to be introduced in the sorting engineering to further influence the sorting result; in addition, in the schemes such as embodiment 2 and embodiment 3, the size of the inlet of the sorting flow channel can be adjusted by controlling the pressure of the pressure pipeline so as to adapt to the selection requirements of different sorting targets, and the requirement of manufacturing precision can be reduced to a certain extent.
The embodiments of the present invention have been described in detail with reference to the accompanying drawings, but the present invention is not limited to the above embodiments, and various changes can be made within the knowledge of those skilled in the art without departing from the gist of the present invention. Furthermore, the embodiments of the present invention and the features of the embodiments may be combined with each other without conflict.

Claims (9)

1. A microfluidic chip comprising a channel layer, the channel layer comprising:
the main flow passage is used for passing the sample liquid;
the separation flow channel is communicated with the main flow channel through a separation flow channel inlet, and the height of the separation flow channel inlet is smaller than that of the main flow channel.
2. The microfluidic chip according to claim 1, wherein the main channel is connected to the sorting channel at both sides in an axial direction thereof through the sorting channel inlet.
3. A microfluidic chip, comprising:
the channel layer comprises a main runner for sample liquid to pass through and a separation runner, and the separation runner is communicated with the main runner through a separation runner inlet;
the control layer is arranged on at least one side of the channel layer in a stacked mode, a pressure pipeline is arranged on the control layer at a position corresponding to the sorting flow channel inlet and used for driving the sorting flow channel inlet to shrink in the height direction, and therefore the height of the sorting flow channel inlet is smaller than that of the main flow channel.
4. The microfluidic chip according to claim 3, wherein a thin film layer is further disposed between the control layer and the channel layer, and the pressure pipeline drives the thin film layer to deform in a height direction of the sorting channel inlet so as to adjust the height of the sorting channel inlet.
5. The microfluidic chip according to claim 4, wherein the pressure channel is inclined to a flow direction of the main channel.
6. The microfluidic chip according to any one of claims 3 to 5, wherein the sorting flow channel is connected to the main flow channel at both sides of the main flow channel in the axial direction thereof through the sorting flow channel inlet.
7. The microfluidic chip according to claim 6, wherein the sorting channel inlets on both sides of the main channel are staggered.
8. The microfluidic chip according to any one of claims 3 to 5, wherein the main channel is further provided with a filtering mechanism.
9. A sorting method using the microfluidic chip according to any one of claims 3 to 8, comprising the steps of:
s1: driving the inlet of the sorting flow channel to contract in the height direction;
s2: inputting sample liquid into the main flow channel;
s3: and respectively collecting sample liquid at the outlet of the main runner and the outlet of the sorting runner.
CN201911240815.6A 2019-12-06 2019-12-06 Microfluidic chip and sorting method Pending CN111040938A (en)

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CN112844502A (en) * 2020-12-30 2021-05-28 厦门天马微电子有限公司 Microfluidic device and manufacturing method and extraction method thereof

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