CN115786074A - Microfluidic chip and method for high-flux rapid and accurate cell sorting at low flow rate - Google Patents
Microfluidic chip and method for high-flux rapid and accurate cell sorting at low flow rate Download PDFInfo
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Abstract
The invention discloses a micro-fluidic chip for high-flux rapid and accurate cell sorting at low flow rate, which comprises an upper cover plate, a flow guide layer, a flow channel layer and a lower cover plate which are sequentially arranged from top to bottom, wherein the flow channel layer comprises a sample flow main inlet flow channel, a sorting unit and a second cell outlet flow channel, and the sorting unit comprises a sample flow double-division inlet flow channel, an extrusion direct flow channel, a contraction and expansion array flow channel and a sudden expansion sorting flow channel which are sequentially connected; the sample flow double-division inlet flow channel comprises a first sample flow inlet flow channel, a sheath flow inlet flow channel and a second sample flow inlet flow channel, and the sudden-expansion separation flow channel comprises a first outlet flow channel, a second outlet flow channel and a third outlet flow channel. The invention realizes the rapid and accurate sorting of the cells, and the detection efficiency of the rare cells is high.
Description
Technical Field
The invention relates to a micro-fluidic chip and a method for realizing rapid and accurate cell sorting based on size by a viscoelastic fluid focusing technology and a contraction and expansion array flow channel sorting technology, belonging to the field of biological particle control of micro-fluidic chips.
Background
Rare cells in blood such as Circulating Tumor Cells (CTCs) have important clinical value, and detection of the circulating tumor cells is beneficial to early diagnosis, chemotherapy, curative effect evaluation and the like of malignant tumors. However, since the content of rare cells is very low, a rare cell detection device which is simple to operate, high in throughput and rapid and accurate becomes a research hotspot.
The microfluidic cell control mode mainly comprises an active mode and a passive mode, the active mode is based on external field acting forces such as sound, magnetism and electricity, the microfluidic cell control method has the advantages of high universality, high accuracy and the like, the passive mode does not need the auxiliary action of an external field, and the microfluidic cell control method has the advantages of high flux, low cost, easiness in operation and the like and has a better application prospect. At present, the Newtonian fluid inertial microfluidic technology is often adopted to improve the sorting efficiency of particles, but the control precision of small-size cells is limited because the sorting is carried out at a higher flow velocity. Viscoelastic fluid has better differentiation precision to not unidimensional particle, nevertheless viscoelastic fluid's work flux is not high, the cell focusing can be accelerated to shrink expansion array, the focusing rate difference of the different size cells of aggravation, realize the quick accurate sorting in the short distance runner, and this runner is convenient for parallel arrangement, the runner flux is promoted at double, promote the detection efficiency of rare cell by a wide margin, so utilize viscoelastic fluid to have very important realistic meaning to the quick accurate sorting effect of rare cell in parallel shrink expansion array runner.
Disclosure of Invention
The purpose of the invention is as follows: in order to overcome the defects in the prior art, the invention provides the microfluidic chip and the method for quickly and accurately sorting the cells at high flux under low flow speed, which can realize high-flux and quick and accurate sorting of the cells.
The technical scheme is as follows: in order to achieve the purpose, the invention adopts the technical scheme that:
the utility model provides a micro-fluidic chip that quick accurate separation of cell high flux under low flow rate, includes by last upper cover plate, water conservancy diversion layer, runner layer and the lower apron that sets gradually under to, wherein:
the flow channel layer comprises a sample flow main inlet flow channel, a sorting unit and a second cell outlet flow channel, wherein one end of the sample flow main inlet flow channel is provided with a sample flow channel layer inlet, and the other end of the sample flow main inlet flow channel is provided with a sample flow inlet. And one end of the second cell outlet flow channel is provided with a flow channel outlet, and the other end of the second cell outlet flow channel is provided with a flow channel layer cell outlet.
The sorting unit comprises a sample flow double-branch inlet flow channel, an extrusion direct flow channel, a contraction and expansion array flow channel and a sudden expansion sorting flow channel which are sequentially connected.
The sample flow double-split inlet flow channel comprises a first sample flow inlet flow channel, a sheath flow inlet flow channel and a second sample flow inlet flow channel, wherein the inlet end of the first sample flow inlet flow channel and the inlet end of the second sample flow inlet flow channel are communicated with a sample flow inlet, and the outlet end of the first sample flow inlet flow channel, the outlet end of the sheath flow inlet flow channel and the outlet end of the second sample flow inlet flow channel are communicated with the inlet end of the extrusion direct flow channel. The inlet end of the sheath inflow port flow channel is provided with a sheath inflow port, the sheath inflow port flow channel is positioned between the first sample inflow port flow channel and the second sample inflow port flow channel, and the first sample inflow port flow channel and the second sample inflow port flow channel are symmetrical relative to the sheath inflow port flow channel.
The sudden-expansion separation flow channel comprises a first outlet flow channel, a second outlet flow channel and a third outlet flow channel, wherein the inlet end of the first outlet flow channel, the inlet end of the second outlet flow channel and the inlet end of the third outlet flow channel are connected with the outlet end of the contraction and expansion array flow channel, and the outlet end of the first outlet flow channel and the outlet end of the third outlet flow channel are connected with the flow channel outlet. The outlet end of the second outlet flow passage is provided with a second outlet flow passage outlet, the second outlet flow passage is positioned between the first outlet flow passage and the third outlet flow passage, and the first outlet flow passage and the third outlet flow passage are symmetrical relative to the second outlet flow passage.
Preferably: the flow guide layer is provided with a sample flow guide layer inlet, a sheath flow guide layer inlet flow channel, a first cell outlet flow channel and a flow guide layer second outlet. One end of the sheath flow guiding layer inlet flow channel is provided with a flow guiding layer sheath flow inlet, the other end of the sheath flow guiding layer inlet flow channel is provided with a sheath flow branch inlet, one end of the first cell outlet flow channel is provided with a flow channel branch outlet, and the other end of the first cell outlet flow channel is provided with a flow guiding layer first outlet.
The sample flow guide layer inlet and the flow channel layer sample flow inlet are communicated from top to bottom in sequence. The sheath flow branch inlets are communicated with the sheath flow inlets from top to bottom in sequence. The flow channel branch outlet and the second outlet flow channel outlet are communicated in sequence from top to bottom. And the second outlet of the flow guide layer and the cell outlet of the flow channel layer are sequentially communicated from top to bottom.
Preferably: the upper cover plate is provided with a sample flow main inlet, a sheath flow main inlet, a first main outlet and a second main outlet. The sample flow total inlet, the sample flow guide layer inlet and the flow passage layer sample flow inlet are communicated from top to bottom in sequence. The sheath flow main inlet and the sheath flow inlet of the diversion layer are sequentially communicated from top to bottom, and the sheath flow branch inlet and the sheath flow inlet are sequentially communicated from top to bottom. The first main outlet and the first outlet of the diversion layer are communicated in sequence from top to bottom. The second total outlet, the second outlet of the diversion layer and the cell outlet of the flow channel layer are communicated in sequence from top to bottom.
Preferably, the following components: the sample inflow port, the sheath inflow port flow channel, the extrusion direct flow channel, the contraction flow channel of the contraction and expansion flow channel, the sudden expansion sorting flow channel, the second outlet flow channel and the flow channel outlet are all positioned on the same axis. The first sample inlet flow channel and the first outlet flow channel are on the same side of the axis, and the second sample inlet flow channel and the third outlet flow channel are on the other side of the axis.
Preferably, the following components: the contraction and expansion array flow passage is formed by alternately arranging and connecting a plurality of expansion flow passages and contraction flow passages, and the adjacent expansion flow passages are communicated with the contraction flow passages.
Preferably, the following components: the contraction and expansion array flow passage is of an axisymmetric structure.
Preferably, the following components: and the included angle between the outlet section of the sheath inflow port flow channel and the outlet section of the first sample inflow port flow channel and the included angle between the outlet section of the sheath inflow port flow channel and the outlet section of the second sample inflow port flow channel are acute angles. And an included angle between the outlet section of the second outlet flow channel and the inlet section of the first outlet flow channel and an included angle between the outlet section of the second outlet flow channel and the inlet section of the third outlet flow channel are acute angles.
Preferably: the number of the sorting units is 4, and the 4 sorting units are arranged in parallel.
A method for quickly and accurately sorting cells at a high flux at a low flow rate adopts the microfluidic chip for quickly and accurately sorting the cells at the high flux at the low flow rate, and comprises the following steps:
step 1, injecting viscoelastic sample flow with mixed blood from a sample main inlet of an upper cover plate, flowing into a sample inlet of a sample flow main inlet flow channel of a flow channel layer through a diversion layer, injecting cell-free viscoelastic sheath flow from a sheath flow main inlet of the upper cover plate, and flowing into a sheath flow inlet of the flow channel layer through a sheath flow inlet flow channel of the flow channel layer.
And 2, extruding the viscoelastic sample flow by the elastic force pointing to the wall surface before entering the contraction and expansion array flow channel due to the action of viscoelastic sheath flow, and moving along two side wall surfaces of the extrusion direct flow channel all the time.
And 3, allowing the cells to enter the contracted and expanded array flow channel and migrate to the center of the flow channel under the action of elastic force of viscoelastic fluid, wherein the elastic force borne by large particles is larger, the cells are subjected to drag force generated by the flow channel at the outlet of each expanded flow channel and migrate to the center of the flow channel, the size of the drag force borne by the cells is in direct proportion to the size of the cells, namely the drag force and the elastic force borne by rare cells are larger, and the migration speed to the position near the center of the flow channel is higher.
And 4, step 4: after flowing through the contraction and expansion array flow channel, the rare cells are positioned near the center of the flow channel, the blood cells are still positioned near the wall surface, after the cells enter the sudden expansion sorting flow channel, the distance between the rare cells and the blood cells is further expanded in the sudden expansion sorting flow channel, the rare cells flow into a first total outlet of the upper cover plate from a second outlet flow channel outlet through a first cell outlet flow channel of the diversion layer, the blood cells on two sides flow into a second total outlet (20) of the upper cover plate (1) from the flow channel outlet through a first outlet flow channel and a third outlet flow channel respectively, and the rapid and accurate sorting of the rare cells (35) and the blood cells (36) in a short distance is realized.
Compared with the prior art, the invention has the following beneficial effects:
the viscoelastic sample flow is extruded to the two sides of the flow channel after entering the extrusion straight flow channel under the action of the viscoelastic sheath flow, cells enter the contraction and expansion flow channel and migrate to the center of the flow channel under the action of the elastic force of the viscoelastic fluid, and large particles are subjected to larger elastic force and have higher focusing speed. The cells are migrated to the center of the flow channel by the drag force generated by the flow channel at the outlet of each expansion flow channel, and the size of the drag force borne by the cells is in direct proportion to the size of the cells, namely, the drag force borne by the rare cells with larger size is larger than that borne by the red cells with smaller size, the traversing speed to the center of the flow channel is higher, and the migration speed difference of the cells with different sizes is increased. After the rare cells flow through the contraction and expansion array, the rare cells are positioned near the center of the flow channel, and the blood cells are still positioned near the wall surface, after the cells enter the sudden expansion sorting flow channel, the distance between the rare cells and the blood cells is further expanded in the sudden expansion sorting flow channel, so that the rapid and accurate sorting of the rare cells and the blood cells is realized. The invention has simple structure, is formed by arranging a plurality of sorting units in parallel, which are manufactured by adopting viscoelastic fluid cell focusing technology and contraction-expansion array flow channel sorting technology, has multiplied processing flux, shortens the length of the required sorting flow channel, greatly improves the sorting efficiency and realizes the rapid and accurate sorting of blood cells.
Drawings
FIG. 1 is an exploded view of the assembly of the structure of the present invention;
FIG. 2 is a top view of the present invention;
FIG. 3 is a schematic view of the upper cover plate structure;
FIG. 4 is a first schematic view of a flow guiding layer structure;
FIG. 5 is a schematic view of a flow guiding layer structure II;
FIG. 6 is a schematic view of a flow channel layer structure;
FIG. 7 is a schematic diagram of the configuration of the sorting unit;
FIG. 8 is a schematic view of the structure of the lower cover plate;
FIG. 9 is a schematic view of the flow channel structure of the contraction and expansion array and the cell separation principle of the present invention;
FIG. 10 is a schematic diagram of the cell sorting achieved by the burst expansion structure of the present invention;
in the figure: 1. upper cover plate, 2, flow guide layer, 3, flow channel layer, 4, lower cover plate, 10, flow channel layer of sorting unit, 11, sample flow double entry flow channel, 111, sample entry, 112, first sample entry flow channel, 113, second sample entry flow channel, 12, sheath entry flow channel, 121, sheath entry flow port, 13, extruded straight flow channel, 14, convergent-divergent array flow channel, 15, sudden-divergent sorting flow channel, 151, first exit flow channel, 152, second exit flow channel, 1521, second exit flow channel exit, 153, third exit flow channel, 16, flow channel exit, 17, sample flow total entry, 18, sheath flow total entry, 19, first total exit, 20, second total exit, 21, sample flow guide layer entry, 22 sheath flow guide layer entry, 23, sheath flow guide layer entry, 24, sheath flow branch entry, 25, flow channel branch exit, 26, first cell exit flow channel, 27, flow guide layer first exit, 28, flow guide layer second exit, 29, sample flow layer entry, 30, sample flow guide layer entry, 31, sample flow guide layer entry, 30, cell flow channel, cell exit, cell flow channel, elastic flow channel, cell exit, cell flow channel, 33, cell flow channel, adhesive layer exit, 35, cell flow channel, adhesive layer exit, 33, cell flow channel, 13.
Detailed Description
The present invention is further illustrated in the accompanying drawings and described in the following detailed description, it is to be understood that such examples are included solely for the purposes of illustration and are not intended as a definition of the limits of the invention, since various equivalent modifications of the invention will become apparent to those skilled in the art after reading the present specification, and it is intended to cover all such modifications as fall within the scope of the invention as defined in the appended claims.
A microfluidic chip for high-throughput rapid and accurate cell sorting at low flow rate, as shown in FIGS. 1 to 10, comprises an upper cover plate 1, a flow guide layer 2, a flow channel layer 3 and a lower cover plate 4, which are sequentially arranged from top to bottom, wherein:
as shown in fig. 2 and 3, the upper cover plate 1 is provided with a sample flow inlet 17, a sheath flow inlet 18, a first outlet 19 and a second outlet 20;
as shown in fig. 4 and 5, the flow guiding layer 2 is provided with a sample flow guiding layer inlet 21, a sheath flow guiding layer inlet flow channel 23, a first cell outlet flow channel 26 and a flow guiding layer second outlet 28; one end of the sheath flow guiding layer inlet channel 23 is provided with a flow guiding layer sheath flow inlet 22, the other end is provided with a sheath flow branch inlet 24, one end of the first cell outlet channel 26 is provided with a channel branch outlet 25, and the other end is provided with a flow guiding layer first outlet 27;
as shown in fig. 6 and 7, the flow channel layer 3 includes a sample flow total inlet flow channel 30, a sorting unit 10, and a second cell outlet flow channel 31, the sample flow total inlet flow channel 30 is provided with a sample flow channel layer inlet 29 at one end and a sample flow inlet 111 at the other end; one end of the second cell outlet flow channel 31 is provided with a flow channel outlet 16, and the other end is provided with a flow channel layer cell outlet 32; the sorting units 10 are connected in parallel, the number of the sorting units is 4, and the 4 sorting units are arranged in parallel.
The sorting unit 10 comprises a sample flow double-branch inlet flow channel 11, an extrusion direct flow channel 13, a contraction and expansion array flow channel 14 and a sudden expansion sorting flow channel 15 which are connected in sequence;
the sample flow double inlet channel 11 comprises a first sample flow inlet channel 112 and a second sample flow inlet channel 113, wherein the inlet end of the first sample flow inlet channel 112 and the inlet end of the second sample flow inlet channel 113 are both communicated with the sample flow inlet 111, and the outlet end of the first sample flow inlet channel 112, the outlet end of the second sample flow inlet channel 113 and the outlet end of the sheath flow inlet channel 12 are all communicated with the inlet end of the extrusion straight channel 13;
the inlet end of the sheath inflow port flow channel 12 is provided with a sheath inflow port 121, the sheath inflow port flow channel 12 is located between the first sample inflow port flow channel 112 and the second sample inflow port flow channel 113, the first sample inflow port flow channel 112 and the second sample inflow port flow channel 113 are symmetrical with respect to the sheath inflow port flow channel 12, and the included angle between the outlet section of the sheath inflow port flow channel 12 and the outlet section of the first sample inflow port flow channel 112 and the outlet section of the second sample inflow port flow channel 113 is an acute angle;
the contraction and expansion array flow passage 14 is formed by alternately arranging and connecting a plurality of expansion flow passages and contraction flow passages, the adjacent expansion flow passages are communicated with the contraction flow passages, and the contraction and expansion array flow passage 14 is in an axisymmetric structure.
The sudden expansion sorting flow channel 15 comprises a first outlet flow channel 151, a second outlet flow channel 152 and a third outlet flow channel 153, wherein the inlet ends of the first outlet flow channel 151, the second outlet flow channel 152 and the third outlet flow channel 153 are all connected with the outlet ends of the contraction and expansion array flow channels 14, and the outlet ends of the first outlet flow channel 151 and the third outlet flow channel 153 are all connected with the flow channel outlet 16;
a second outlet flow channel outlet 1521 is arranged at the outlet end of the second outlet flow channel 152, the second outlet flow channel 152 is located between the first outlet flow channel 151 and the third outlet flow channel 153, the first outlet flow channel 151 and the third outlet flow channel 153 are symmetrical with respect to the second outlet flow channel 152, and an acute angle is formed between the outlet section of the second outlet flow channel 152 and the inlet section of the first outlet flow channel 151 and the inlet section of the third outlet flow channel 153;
the sample inflow port 111, the sheath inflow port 121, the sheath inflow port flow channel 12, the extrusion straight flow channel 13, the contraction flow channel of the contraction and expansion flow channel 14, the sudden expansion sorting flow channel 15, the second outlet flow channel 152 and the flow channel outlet 16 are all positioned on the same axis; the first sample inlet channel 112 and the first outlet channel 151 are on the same side of the axis, and the second sample inlet channel 113 and the third outlet channel 153 are on the other side of the axis;
the sample flow total inlet 17, the sample flow guide layer inlet 21 and the flow passage layer sample flow inlet 29 are communicated from top to bottom in sequence; the sheath flow main inlet 18 and the sheath flow inlet 22 of the flow guide layer are communicated with each other from top to bottom, and the sheath flow branch inlets 24 are communicated with the sheath flow inlet 121 from top to bottom; the first main outlet 19 and the first outlet 27 of the flow guide layer are sequentially communicated from top to bottom, and the flow channel branch outlet 25 and the second outlet flow channel outlet 1521 are sequentially communicated from top to bottom; the second total outlet 20, the diversion layer second outlet 28 and the runner layer cell outlet 32 are communicated in sequence from top to bottom;
the sample flow main inlet 17 and the sheath flow main inlet 18 of the upper cover plate 1 are respectively filled with a mixed cell viscoelastic sample flow 33 and a cell-free viscoelastic sheath flow 34; the viscoelastic sample flow 33 and the viscoelastic sheath flow 34 are viscoelastic solutions, including polyvinylpyrrolidone solutions. The flow rate of the solution in the sample flow dual inlet channel 11 is the same as the flow rate of the solution in the sheath flow inlet channel 12, and the reynolds number is equal to about 1.
The mixed large-size cell solution flows into the first cell outlet flow channel 26 of the flow guiding layer 2 through the second outlet flow channel outlet 1521 and then flows into the first total outlet 19 of the upper cover plate 1, and the mixed small-size cell solution flows into the flow channel outlet 16 through the first outlet flow channel 151 and the third outlet flow channel 153 and then flows into the second total outlet 20 of the upper cover plate 1 through the second cell outlet flow channel 31.
The flow guide layer is provided with a sample flow guide layer inlet, a sheath flow guide layer inlet flow channel, a first cell outlet flow channel and a second flow guide outlet, the flow channel layer is provided with a sample flow main inlet flow channel, a sorting unit and a second cell outlet flow channel, and the flow channel layer is formed by parallelly arranging and connecting a plurality of sorting units. The sorting unit is provided with a sample flow double-branch inlet flow channel, a sheath flow inlet flow channel, an extrusion straight flow channel, a contraction and expansion array flow channel and a sudden expansion sorting flow channel. Sheath flow and sample flow are viscoelastic solution, have better differentiation precision to different size cells, and the cell all receives the elastic force effect to flow channel center migration in viscoelastic fluid, and the focusing speed of different size particles is different, thereby the shrink expansion array accelerates particle focusing and aggravates the quick accurate sorting of particle focusing speed difference realization cell.
A method for rapidly and accurately sorting cells at a low flow rate in a high flux manner is disclosed, as shown in FIGS. 9 and 10, the microfluidic chip for rapidly and accurately sorting cells at a low flow rate in a high flux manner comprises the following steps:
step 1: injecting a viscoelastic sample flow 33 with mixed blood from the sample inlet 17 of the upper cover plate 1, flowing into the sample inlet 30 of the channel layer 3 through the flow guiding layer 2, and then flowing into the sample inlet 111, and injecting a cell-free viscoelastic sheath flow 34 from the sheath inlet 18 of the upper cover plate 1, and flowing into the sheath inlet 121 of the channel layer 3 through the sheath flow guiding layer inlet channel 23 of the flow guiding layer 2;
step 2: the viscoelastic sample flow 33 is pressed by the elastic force directed to the wall surface by the viscoelastic sheath flow 34 before entering the flow channel of the contracted/expanded array, and moves along both side wall surfaces of the pressing straight flow channel 13.
And 3, the cells enter the contracted and expanded array flow channel 14 and migrate to the center of the flow channel under the action of the elastic force of the viscoelastic fluid, the elastic force borne by large particles is larger, the cells migrate to the center of the flow channel at the outlet of each expanded flow channel under the drag force generated by the flow channel, the size of the drag force borne by the cells is in direct proportion to the size of the cells, namely the drag force and the elastic force borne by the rare cells 35 are larger, and the migration speed to the vicinity of the center of the flow channel is higher.
As shown in FIG. 9, rare cells and 35 blood cells 36 enter the flow channel 14 along the wall surface in a contracted-expanded configuration where the cells are under the elastic force F of the viscoelastic fluid E The elastic force F borne by the large particles is transferred to the center of the flow passage under the action of the elastic force F E Larger, and generates a drag force F towards the center of the flow channel on the cell at the outlet of each contraction and expansion structure D Pushing the cells to the center of the flow channel with a drag force F D Is proportional to the cell size, i.e., the larger size rare cell 35 experiences a drag force F on the smaller size blood cell 36 D And elastic force F E Larger, rare cells 35 have a faster lateral migration rate, so after a series of contraction-expansion arrays, the rare cells 35 are near the center of the flow channel, while the blood cells 36 remain near the wall.
And 4, step 4: after flowing through the contraction and expansion array flow channel, the rare cell 35 is already positioned near the center of the flow channel, and the blood cell 36 is still positioned near the wall surface, after the cell enters the sudden expansion sorting flow channel 15, the distance between the rare cell 35 and the blood cell 36 is further expanded in the sudden expansion sorting flow channel 15, the rare cell 35 flows into the first total outlet 19 of the upper cover plate 1 from the second outlet flow channel 1521 through the first cell outlet flow channel 23 of the flow guiding layer 2, the blood cells 30 on both sides flow into the flow channel outlet 16 through the first outlet flow channel 151 and the third outlet flow channel 153, and then flow into the second total outlet 20 through the second cell outlet flow channel 31, so that the rapid and accurate sorting of the rare cell 35 and the blood cell 36 in a short distance is realized, and the sorting units 10 are arranged in parallel, and the flow channel flux is multiplied.
As shown in fig. 10, due to the widening of the flow channel, the distance between the rare cell 35 and the blood cell 36 will be further increased, the rare cell 35 flows from the second outlet flow channel 1521 through the first cell outlet flow channel 23 of the diversion layer 2 into the first total outlet 19 of the upper cover plate 1, the blood cells 30 on both sides flow into the flow channel outlet 16 through the first outlet flow channel 151 and the third outlet flow channel 153, and then flow into the second total outlet 20 through the second cell outlet flow channel 31, so as to realize the rapid and accurate sorting of the rare cell 35 and the blood cell 36 in a short distance, and since the sorting units 10 are arranged in parallel, the flow channel flux is increased by times, and the detection efficiency of the rare cell 35 is greatly improved.
In the present invention, a viscoelastic solution is used for both the viscoelastic sheath fluid 33 and the viscoelastic sample fluid 34, and the flow rate of the viscoelastic sample fluid 33 in the sample fluid dual inlet channel 11 is consistent with the flow rate of the viscoelastic sheath fluid 34 in the sheath fluid inlet channel 12. The speed that the cell that the size is great is more fast than the less cell of size to runner center sideslip, has realized the quick accurate sorting in simple runner structure, because the runner sets up in parallel, promotes runner flux at double, promotes rare cell's detection efficiency by a wide margin.
The above description is only of the preferred embodiments of the present invention, and it should be noted that: it will be apparent to those skilled in the art that various modifications and adaptations can be made without departing from the principles of the invention and these are intended to be within the scope of the invention.
Claims (9)
1. The utility model provides a micro-fluidic chip that quick accurate sorting of cell high flux under low velocity of flow which characterized in that: include by upper cover plate (1), water conservancy diversion layer (2), runner layer (3) and apron (4) down that set gradually under to, wherein:
the flow channel layer (3) comprises a sample flow total inlet flow channel (30), a sorting unit (10) and a second cell outlet flow channel (31), one end of the sample flow total inlet flow channel (30) is provided with a sample flow channel layer inlet (29), and the other end is provided with a sample flow inlet (111); one end of the second cell outlet flow channel (31) is provided with a flow channel outlet (16), and the other end is provided with a flow channel layer cell outlet (32);
the sorting unit (10) comprises a sample flow double-branch inlet flow channel (11), an extrusion direct flow channel (13), a contraction and expansion array flow channel (14) and a sudden expansion sorting flow channel (15) which are connected in sequence;
the sample flow double-inlet flow channel (11) comprises a first sample flow inlet flow channel (112), a sheath flow inlet flow channel (12) and a second sample flow inlet flow channel (113), wherein the inlet end of the first sample flow inlet flow channel (112) and the inlet end of the second sample flow inlet flow channel (113) are communicated with a sample flow inlet (111), and the outlet end of the first sample flow inlet flow channel (112), the outlet end of the second sample flow inlet flow channel (113) and the outlet end of the sheath flow inlet flow channel (12) are communicated with the inlet end of the extrusion straight flow channel (13); the inlet end of the sheath inflow port flow channel (12) is provided with a sheath inflow port (121), the sheath inflow port flow channel (12) is positioned between the first sample inflow port flow channel (112) and the second sample inflow port flow channel (113), and the first sample inflow port flow channel (112) and the second sample inflow port flow channel (113) are symmetrical relative to the sheath inflow port flow channel (12);
the sudden-expansion sorting flow channel (15) comprises a first outlet flow channel (151), a second outlet flow channel (152) and a third outlet flow channel (153), the inlet end of the first outlet flow channel (151), the inlet end of the second outlet flow channel (152) and the inlet end of the third outlet flow channel (153) are connected with the outlet end of the contraction and expansion array flow channel (14), and the outlet end of the first outlet flow channel (151) and the outlet end of the third outlet flow channel (153) are connected with a flow channel outlet (16); the outlet end of the second outlet flow channel (152) is provided with a second outlet flow channel outlet (1521), the second outlet flow channel (152) is positioned between the first outlet flow channel (151) and the third outlet flow channel (153), and the first outlet flow channel (151) and the third outlet flow channel (153) are symmetrical relative to the second outlet flow channel (152).
2. The microfluidic chip for high-throughput rapid and accurate sorting of cells at low flow rate according to claim 1, wherein: the flow guide layer (2) is provided with a sample flow guide layer inlet (21), a sheath flow guide layer inlet flow channel (23), a first cell outlet flow channel (26) and a flow guide layer second outlet (28); one end of the sheath flow guiding layer inlet flow passage (23) is provided with a flow guiding layer sheath flow inlet (22), the other end of the sheath flow guiding layer inlet flow passage is provided with a sheath flow branch inlet (24), one end of the first cell outlet flow passage (26) is provided with a flow passage branch outlet (25), and the other end of the first cell outlet flow passage is provided with a flow guiding layer first outlet (27);
the sample flow guide layer inlet (21) and the flow passage layer sample flow inlet (29) are communicated in sequence from top to bottom; the sheath flow branch inlet (24) is communicated with the sheath flow inlet (121) from top to bottom in sequence; the flow channel branch outlet (25) and the second outlet flow channel outlet (1521) are communicated in sequence from top to bottom; the second outlet (28) of the flow guide layer and the cell outlet (32) of the flow channel layer are sequentially communicated from top to bottom.
3. The microfluidic chip for high-throughput rapid and accurate cell sorting at low flow rate according to claim 2, wherein: the upper cover plate (1) is provided with a sample flow main inlet (17), a sheath flow main inlet (18), a first main outlet (19) and a second main outlet (20); the sample flow total inlet (17), the sample flow guide layer inlet (21) and the flow passage layer sample flow inlet (29) are communicated from top to bottom in sequence; the sheath flow main inlet (18) and the flow guide layer sheath flow inlet (22) are sequentially communicated from top to bottom, and the sheath flow branch inlet (24) and the sheath flow inlet (121) are sequentially communicated from top to bottom; the first main outlet (19) and the first outlet (27) of the diversion layer are communicated in sequence from top to bottom; the second total outlet (20), the diversion layer second outlet (28) and the flow channel layer cell outlet (32) are communicated from top to bottom in sequence.
4. The microfluidic chip for high-throughput rapid and accurate sorting of cells at low flow rate according to claim 3, wherein: the sample inflow port (111), the sheath inflow port (121), the sheath inflow port flow channel (12), the extrusion direct flow channel (13), the contraction flow channel of the contraction and expansion flow channel (14), the sudden expansion sorting flow channel (15), the second outlet flow channel (152) and the flow channel outlet (16) are all positioned on the same axis; the first sample inlet flow channel (112) is on the same side of the axis as the first outlet flow channel (151) and the second sample inlet flow channel (113) is on the other side of the axis as the third outlet flow channel (153).
5. The microfluidic chip for high-throughput rapid and accurate cell sorting at low flow rate according to claim 4, wherein: the contraction and expansion array flow passage (14) is formed by alternately arranging and connecting a plurality of expansion flow passages and contraction flow passages, and the adjacent expansion flow passages are communicated with the contraction flow passages.
6. The microfluidic chip for high-throughput rapid and accurate cell sorting at low flow rate according to claim 5, wherein: the contraction and expansion array flow passage (14) is of an axisymmetric structure.
7. The microfluidic chip for high-throughput rapid and accurate cell sorting at low flow rate according to claim 6, wherein: an included angle between the outlet section of the sheath inflow port flow channel (12) and the outlet section of the first sample inflow port flow channel (112) and the outlet section of the second sample inflow port flow channel (113) is an acute angle; and an included angle between the outlet section of the second outlet flow channel (152) and the inlet section of the first outlet flow channel (151) and an included angle between the outlet section of the second outlet flow channel and the inlet section of the third outlet flow channel (153) are acute angles.
8. The microfluidic chip for high-throughput rapid and accurate cell sorting at low flow rate according to claim 7, wherein: the number of the sorting units (10) is 4, and the 4 sorting units (10) are arranged in parallel.
9. A high-flux rapid and accurate cell sorting method at low flow rate is characterized in that: the microfluidic chip for high-throughput rapid and accurate sorting of cells at low flow rate according to claim 1, comprising the steps of:
step 1, injecting a viscoelastic sample flow (33) with mixed blood from a sample total inlet (17) of an upper cover plate (1), flowing into a sample inlet (111) of a sample flow total inlet flow channel (30) of a flow channel layer (3) through a diversion layer (2), injecting a cell-free viscoelastic sheath flow (34) from a sheath flow total inlet (18) of the upper cover plate (1), and flowing into a sheath flow inlet (121) of the flow channel layer (3) through a diversion layer sheath flow inlet flow channel (23) of the diversion layer (2);
step 2, the viscoelastic sample flow (33) is extruded by the elastic force pointing to the wall surface before entering the contraction and expansion array flow passage (14) due to the effect of the viscoelastic sheath flow (34), and moves along the two side wall surfaces of the extrusion straight flow passage (13);
step 3, the cells enter the contracted and expanded array flow channel (14) and migrate to the center of the flow channel under the action of elastic force of viscoelastic fluid, the elastic force borne by large particles is larger, the cells migrate to the center of the flow channel under the drag force generated by the flow channel at the outlet of each expanded flow channel, the size of the drag force borne by the cells is in direct proportion to the size of the cells, namely the drag force and the elastic force borne by the rare cells (35) are larger, and the migration speed to the position near the center of the flow channel is higher;
and 4, step 4: after the cells flow through the contraction and expansion array flow channel (14), the rare cells (35) are located near the center of the flow channel, the blood cells (36) are still located near the wall surface, after the cells enter the sudden expansion sorting flow channel (15), the distance between the rare cells (35) and the blood cells (36) is further expanded in the sudden expansion sorting flow channel (15), the rare cells (35) flow from the second outlet flow channel outlet (1521) to the first total outlet (19) of the upper cover plate (1) through the first cell outlet flow channel (26) of the diversion layer (2), and the blood cells (36) on the two sides respectively flow from the flow channel outlet (16) to the second total outlet (20) of the upper cover plate (1) through the second cell outlet flow channel (31) through the first outlet flow channel (151) and the third outlet flow channel (153), so that the rapid rare and accurate sorting of the cells (35) and the blood cells (36) in a short distance is realized.
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