CN115786074B - Microfluidic chip and method for high-throughput rapid and accurate cell sorting at low flow rate - Google Patents
Microfluidic chip and method for high-throughput rapid and accurate cell sorting at low flow rate Download PDFInfo
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Abstract
The application 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 total inlet flow channel, a sorting unit and a second cell outlet flow channel, and the sorting unit comprises a sample flow double-split inlet flow channel, an extrusion straight flow channel, a shrinkage 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 inflow flow channel, a sheath inflow flow channel and a second sample inflow flow channel, and the sudden expansion sorting flow channel comprises a first outlet flow channel, a second outlet flow channel and a third outlet flow channel. The application realizes rapid and accurate cell sorting, and has high detection efficiency of rare cells.
Description
Technical Field
The application relates to a micro-fluidic chip and a method for realizing quick and accurate cell sorting based on size by using a viscoelastic fluid focusing technology and a shrinkage 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, efficacy evaluation and the like of malignant tumors. However, the rare cell detection device which is simple to operate, high in flux and rapid and accurate is a research hot spot because the content of the rare cells is extremely low.
The micro-fluidic cell control mode is mainly divided into an active mode and a passive mode, wherein the active mode has the advantages of high universality, high accuracy and the like by means of external field acting force such as sound, magnetism, electricity and the like, the passive mode does not need external field auxiliary action, and the micro-fluidic cell control method has the advantages of high flux, low cost, easiness in operation and the like, and has a good application prospect. At present, newton fluid inertial microfluidic technology is often adopted to improve the sorting efficiency of particles, but because the particle sorting is performed at a higher flow rate, the control precision of small-size cells is limited. The viscoelastic fluid has better distinguishing precision for particles with different sizes, but the working flux of the viscoelastic fluid is not high, the contraction and expansion array can accelerate cell focusing, the focusing speed difference of the cells with different sizes is increased, quick and accurate sorting in a short-distance flow channel is realized, the flow channels are convenient to be arranged in parallel, the flow channel flux is improved in multiple, and the detection efficiency of rare cells is greatly improved, so that the use of the viscoelastic fluid has very important practical significance for quick and accurate sorting of the rare cells in the parallel contraction and expansion array flow channels.
Disclosure of Invention
The application aims to: in order to overcome the defects in the prior art, the application provides a micro-fluidic chip and a method for realizing high-flux rapid and accurate cell sorting under the conditions of high-flux, rapid and accurate cell sorting and low flow rate.
The technical scheme is as follows: in order to achieve the above purpose, the application adopts the following technical scheme:
the utility model provides a micro-fluidic chip of quick accurate separation of cell high flux under low velocity of flow, includes upper cover plate, guiding layer, runner layer and the lower apron that from top to bottom set gradually, wherein:
the flow channel layer comprises a sample flow total inlet flow channel, a sorting unit and a second cell outlet flow channel, wherein one end of the sample flow total inlet flow channel is provided with a sample flow channel layer inlet, and the other end of the sample flow total inlet flow channel is provided with a sample flow inlet. One end of the second cell outlet runner is provided with a runner outlet, and the other end of the second cell outlet runner is provided with a runner layer cell outlet.
The sorting unit comprises a sample flow double-split inlet runner, an extrusion straight runner, a shrinkage expansion array runner and a sudden expansion sorting runner which are connected in sequence.
The sample flow double-split inlet runner comprises a first sample inflow runner, a sheath inflow runner and a second sample inflow runner, wherein the inlet end of the first sample inflow runner and the inlet end of the second sample inflow runner are communicated with the sample inflow, and the outlet end of the first sample inflow runner, the outlet end of the sheath inflow runner and the outlet end of the second sample inflow runner are communicated with the inlet end of the extrusion straight runner. The inlet end of the sheath inflow port runner is provided with a sheath inflow port, the sheath inflow port runner is positioned between the first sample inflow port runner and the second sample inflow port runner, and the first sample inflow port runner and the second sample inflow port runner are symmetrical with respect to the sheath inflow port runner.
The sudden expansion sorting 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 all connected with the outlet end of the shrinkage 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 all connected with the flow channel outlet. The outlet end of the second outlet runner is provided with a second outlet runner outlet, the second outlet runner is positioned between the first outlet runner and the third outlet runner, and the first outlet runner and the third outlet runner are symmetrical with respect to the second outlet runner.
Preferably: the flow guiding layer is provided with a sample flow guiding layer inlet, a sheath flow guiding layer inlet flow channel, a first cell outlet flow channel and a guiding layer second outlet. One end of the sheath flow diversion layer inlet runner is provided with a diversion layer sheath flow inlet, the other end is provided with a sheath flow branch inlet, one end of the first cell outlet runner is provided with a runner branch outlet, and the other end of the first cell outlet runner is provided with a diversion layer first outlet.
The inlet of the sample flow guide layer and the inlet of the sample flow channel layer are sequentially communicated from top to bottom. The sheath flow branch inlet and the sheath flow inlet are sequentially communicated from top to bottom. The branch outlet of the flow passage and the outlet of the second outlet flow passage are sequentially communicated from top to bottom. The second outlet of the diversion 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 total inlet, a sheath flow total inlet, a first total outlet and a second total outlet. The sample flow total inlet, the sample flow diversion layer inlet and the sample flow passage layer inlet are sequentially communicated from top to bottom. The sheath flow main inlet and the diversion layer sheath flow inlet 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 total outlet and the first outlet of the diversion layer are sequentially communicated 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 sequentially communicated from top to bottom.
Preferably: the sample inflow port, the sheath inflow port flow channel, the extrusion straight flow channel, the shrinkage flow channel of the shrinkage expansion array flow channel, the sudden expansion separation 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 positioned on the same side of the axis, and the second sample inlet flow channel and the third outlet flow channel are positioned on the other side of the axis.
Preferably: the shrinkage expansion array flow passage is formed by alternately arranging and connecting a plurality of expansion flow passages and shrinkage flow passages, and the adjacent expansion flow passages and the shrinkage flow passages are mutually communicated.
Preferably: the shrinkage and expansion array flow passage is of an axisymmetric structure.
Preferably: the included angle between the outlet section of the sheath inflow port runner and the outlet section of the first sample inflow port runner and the outlet section of the second sample inflow port runner is an acute angle. The included angle between the outlet section of the second outlet flow channel and the inlet section of the first outlet flow channel and the inlet section of the third outlet flow channel are acute angles respectively.
Preferably: the number of the sorting units is 4, and the 4 sorting units are arranged in parallel.
A high-flux rapid and accurate cell sorting method under a low flow rate adopts the micro-fluidic chip for high-flux rapid and accurate cell sorting under the low flow rate, and comprises the following steps:
step 1, injecting a viscoelastic sample flow with mixed blood from a sample flow main inlet of an upper cover plate, flowing into a sample flow inlet of a sample flow main inlet runner of a runner layer through a diversion layer, injecting a cell-free viscoelastic sheath flow from a sheath flow main inlet of the upper cover plate, and flowing into a sheath flow inlet of a runner layer through a sheath flow diversion layer inlet runner of the diversion layer.
Step 2, the viscoelastic sample flow is extruded by elastic force directed to the wall surface before entering the shrinkage and expansion array flow channel due to the action of the viscoelastic sheath flow, and moves along the two side wall surfaces of the extrusion direct flow channel.
And 3, allowing the cells to enter the convergent-divergent array flow channel to migrate towards the center of the flow channel under the action of the elastic force of viscoelastic fluid, wherein the elastic force borne by the large particles is larger, the cells migrate towards the center of the flow channel under the action of the drag force generated by the flow channel at the outlet of each divergent 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 and the elastic force borne by rare cells are larger, and the migration speed to the vicinity of the center of the flow channel is higher.
Step 4: after flowing through the shrinkage and expansion array runner, rare cells are positioned near the center of the runner and blood cells are still positioned near the wall surface, after the cells enter the sudden expansion sorting runner, the distance between the rare cells and the blood cells is further enlarged in the sudden expansion sorting runner, the rare cells flow into a first total outlet of an upper cover plate from an outlet of a second outlet runner through a first cell outlet runner of a flow guiding layer, and the blood cells at two sides flow into a second total outlet of the upper cover plate from an outlet of the runner through a second cell outlet runner through a first outlet runner and a third outlet runner respectively, so that the rapid and accurate sorting of the rare cells and the blood cells in a short distance is realized.
Compared with the prior art, the application has the following beneficial effects:
according to the application, the viscoelastic sample flow is extruded to two sides of the flow channel after entering the extrusion straight flow channel due to the action of the viscoelastic sheath flow, cells enter the shrinkage and expansion array flow channel and migrate towards the center of the flow channel under the action of the elastic force of the viscoelastic fluid, and the 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 exerted by the cells is in direct proportion to the cell size, namely, the drag force exerted by rare cells with larger sizes is larger than that exerted by red cells with smaller sizes, so that the speed of traversing to the center of the flow channel is faster, and the migration speed difference of the cells with different sizes is increased. After flowing 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, and after the cells enter the sudden expansion separation flow channel, the distance between the rare cells and the blood cells is further enlarged in the sudden expansion separation flow channel, so that the rapid and accurate separation of the rare cells and the blood cells is realized. The application has simple structure, and is formed by arranging a plurality of sorting units in parallel, wherein the sorting units are manufactured by adopting a viscoelastic fluid cell focusing technology and a shrinkage and expansion array flow channel sorting technology, the processing flux is multiplied, the length of a required sorting flow channel is shortened, the sorting efficiency is greatly improved, and the rapid and accurate sorting of blood cells is realized.
Drawings
Fig. 1 is an exploded view of an assembly of the structure of the present application.
Fig. 2 is a top view of the present application.
Fig. 3 is a schematic view of the structure of the upper cover plate.
Fig. 4 is a schematic diagram of a diversion layer structure.
Fig. 5 is a schematic diagram of a second flow guiding layer structure.
Fig. 6 is a schematic view of a flow channel layer structure.
Fig. 7 is a schematic diagram of the structure of the sorting unit.
Fig. 8 is a schematic structural view of the lower cover plate.
FIG. 9 is a schematic diagram of the flow channel structure of the systolic array and the cell separation principle of the present application.
FIG. 10 is a schematic diagram showing cell sorting achieved by the inventive flash-expanded structure.
In the figure: 1. an upper cover plate, 2, a flow guiding layer, 3, a flow channel layer, 4, a lower cover plate, 10, a sorting unit, 11, a sample flow double-split inlet flow channel, 111, a sample inflow port, 112, a first sample inflow port flow channel, 113, a second sample inflow port flow channel, 12, a sheath inflow port flow channel, 121, a sheath inflow port, 13, an extrusion straight flow channel, 14, a shrinkage expansion array flow channel, 15, a burst sorting flow channel, 151, a first outlet flow channel, 152, a second outlet flow channel, 1521, a second outlet flow channel outlet, 153, a third outlet flow channel, 16, a flow channel outlet, 17, a sample flow total inlet, 18, a sheath flow total inlet, 19, a first total outlet, 20, a second total outlet, 21, a sample flow diversion layer inlet, 22, a diversion layer sheath inlet, 23, a sheath flow diversion layer inlet runner, 24, a sheath flow branch inlet, 25, a runner branch outlet, 26, a first cell outlet runner, 27, a diversion layer first outlet, 28, a diversion layer second outlet, 29, a sample flow runner layer inlet, 30, a sample flow total inlet runner, 31, a second cell outlet runner, 32, a runner layer cell outlet, 33, a viscoelastic sample flow, 34, a viscoelastic sheath flow, 35, rare cells, 36, and blood cells.
Description of the embodiments
The present application is further illustrated in the accompanying drawings and detailed description which are to be understood as being merely illustrative of the application and not limiting of its scope, and various equivalent modifications to the application will fall within the scope of the application as defined in the appended claims after reading the application.
The utility model provides a micro-fluidic chip of quick accurate separation of cell high flux under low velocity of flow, as shown in fig. 1-10, includes upper cover plate 1, water conservancy diversion layer 2, runner layer 3 and the lower apron 4 that set gradually from top to bottom, wherein:
as shown in fig. 2 and 3, the upper cover plate 1 is provided with a sample flow total inlet 17, a sheath flow total inlet 18, a first total outlet 19 and a second total outlet 20.
As shown in fig. 4 and 5, the fluidic layer 2 is provided with a sample flow fluidic layer inlet 21, a sheath flow fluidic layer inlet channel 23, a first cell outlet channel 26, and a fluidic layer second outlet 28. The sheath flow diversion layer inlet runner 23 one end is provided with diversion layer sheath flow inlet 22, and the other end is provided with sheath flow branch inlet 24, and first cell outlet runner 26 one end is provided with runner branch export 25, and the other end is provided with diversion layer first export 27.
As shown in fig. 6 and 7, the flow path layer 3 includes a total sample flow inlet flow path 30, a sorting unit 10, and a second cell outlet flow path 31, and the total sample flow inlet flow path 30 is provided with a sample flow path layer inlet 29 at one end and a sample flow inlet 111 at the other end. The second cell outlet channel 31 has a channel outlet 16 at one end and a channel layer cell outlet 32 at the other end. The plurality of 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-split inlet runner 11, an extrusion straight runner 13, a shrinkage expansion array runner 14 and a sudden expansion sorting runner 15 which are connected in sequence.
The sample flow double-split inlet flow channel 11 comprises a first sample flow inlet flow channel 112 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 the 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 flow inlet channel 12 is provided with a sheath flow inlet 121, the sheath flow inlet channel 12 is located between the first sample flow inlet channel 112 and the second sample flow inlet channel 113, the first sample flow inlet channel 112 and the second sample flow inlet channel 113 are symmetrical with respect to the sheath flow inlet channel 12, and the outlet section of the sheath flow inlet channel 12 is respectively at an acute angle with the included angle between the outlet section of the first sample flow inlet channel 112 and the outlet section of the second sample flow inlet channel 113.
The shrinkage-expansion array flow passage 14 is formed by alternately arranging and connecting a plurality of expansion flow passages and shrinkage flow passages, and the adjacent expansion flow passages are mutually communicated with the shrinkage flow passages, and the shrinkage-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 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 all connected with the outlet end of the shrinkage-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 all connected with the 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 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 the outlet section of the second outlet flow channel 152 is respectively acute with the included angle between the inlet section of the first outlet flow channel 151 and the inlet section of the third outlet flow channel 153.
The sample inlet 111, the sheath inlet 121, the sheath inlet channel 12, the extrusion straight channel 13, the convergent channel of the convergent-divergent array channel 14, the convergent channel 15, the second outlet channel 152 and the channel outlet 16 are all on the same axis. The first sample inflow channel 112 is on the same side of the axis as the first outlet channel 151, and the second sample inflow channel 113 is on the other side of the axis as the third outlet channel 153.
The sample flow total inlet 17, the sample flow guide layer inlet 21 and the sample flow channel layer inlet 29 are sequentially communicated from top to bottom. The sheath flow main inlet 18 and the diversion 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 total outlet 19 and the first outlet 27 of the diversion layer are sequentially communicated from top to bottom, and the runner branch outlet 25 and the second outlet runner outlet 1521 are sequentially communicated from top to bottom. The second total outlet 20, the second outlet 28 of the diversion layer and the cell outlet 32 of the runner layer are sequentially communicated from top to bottom.
The sample flow total inlet 17 and the sheath flow total inlet 18 of the upper cover plate 1 are respectively filled with a viscoelastic sample flow 33 of mixed cells and a viscoelastic sheath flow 34 of no cells. The viscoelastic sample stream 33 and the viscoelastic sheath stream 34 are both viscoelastic solutions, including polyvinylpyrrolidone solutions. The flow rate of the solution in the sample flow dual-split inlet flow channel 11 is identical to the flow rate of the solution in the sheath flow inlet flow channel 12, and the Reynolds number is approximately equal to 1.
The mixed large-size cell solution flows into the first cell outlet channel 26 of the guide layer 2 through the second outlet 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 channel outlet 16 through the first outlet channel 151 and the third outlet channel 153 and then flows into the second total outlet 20 of the upper cover plate 1 through the second cell outlet channel 31.
The flow guide layer is provided with a sample flow guide layer inlet, a sheath flow guide layer inlet runner, a first cell outlet runner and a second flow guide outlet, the runner layer is provided with a sample flow main inlet runner, a sorting unit and a second cell outlet runner, and the runner layer is formed by parallel arrangement and connection of a plurality of sorting units. The separation unit is provided with a sample flow double-split inlet runner, a sheath inflow runner, an extrusion straight runner, a shrinkage expansion array runner and a sudden expansion separation runner. The sheath flow and the sample flow are viscoelastic solutions, so that good differentiation precision is realized for cells with different sizes, the cells are all subjected to elastic force in the viscoelastic fluid to migrate towards the center of the flow channel, and the focusing speeds of particles with different sizes are different, so that the focusing speed difference of the particles is increased by contracting and expanding the array to accelerate the focusing of the particles, and the rapid and accurate sorting of the cells is realized.
The method for rapidly and accurately sorting the cells at high flux under the low flow rate is as shown in fig. 9 and 10, and the microfluidic chip for rapidly and accurately sorting the cells at high flux under the low flow rate comprises the following steps:
step 1: the viscoelastic sample flow 33 with mixed blood is injected from the total sample flow inlet 17 of the upper cover plate 1, flows into the total sample flow inlet flow channel 30 of the flow channel layer 3 through the flow guiding layer 2, flows into the sample flow inlet 111, and the cell-free viscoelastic sheath flow 34 is injected from the total sheath flow inlet 18 of the upper cover plate 1, flows into the sheath flow inlet 121 of the flow channel layer 3 through the sheath flow guiding layer inlet flow channel 23 of the flow guiding layer 2.
Step 2: the viscoelastic sample flow 33 is extruded by the elastic force directed to the wall surface before entering the convergent-divergent array flow channel due to the viscoelastic sheath flow 34, and moves along the two side wall surfaces of the extrusion straight flow channel 13.
Step 3, the cells enter the convergent-divergent array runner 14 to migrate towards the center of the runner under the action of the elastic force of the viscoelastic fluid, the elastic force borne by the large particles is larger, the cells migrate towards the center of the runner under the drag force generated by the runner at the outlet of each convergent-divergent array runner, the drag force borne by the cells is proportional to the cell size, 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 runner is higher.
As shown in FIG. 9, rare cells and 35 blood cells 36 enter the systolic array flow channel 14 along the wall, in the systolic configuration, the cells are under the elastic force F of the viscoelastic fluid E Migration to the center of the flow channel under the action of the elastic force F applied by the large particles E Greater and at the exit of each constriction and expansion structure will generate a drag force F on the cells directed towards the centre of the flow channel D Pushing the cells towards the centre of the flow channel, drag force F D Is proportional to the cell size, i.e. the larger rare cells 35 are subjected to a drag force F than the smaller blood cells 36 D And elastic force F E The larger, rare cells 35 have a faster lateral migration velocity, so that after a series of contracting and expanding arrays, the rare cells 35 are near the center of the flow channel while the blood cells 36 remain near the wall.
Step 4: after flowing through the shrinkage-expansion array runner, the rare cells 35 are positioned near the center of the runner and the blood cells 36 are still positioned near the wall surface, after the cells enter the sudden expansion sorting runner 15, the distance between the rare cells 35 and the blood cells 36 is further expanded in the sudden expansion sorting runner 15, the rare cells 35 flow into the first total outlet 19 of the upper cover plate 1 from the second outlet runner outlet 1521 through the first cell outlet runner 26 of the diversion layer 2, the blood cells 36 at two sides flow into the runner outlet 16 through the first outlet runner 151 and the third outlet runner 153 respectively, and then flow into the second total outlet 20 through the second cell outlet runner 31, so that the rapid and accurate sorting of the rare cells 35 and the blood cells 36 in a short distance is realized, and the runner flux is doubly improved due to the parallel arrangement of the sorting units 10.
As shown in fig. 10, due to the widening of the flow channel, the distance between the rare cells 35 and the blood cells 36 is further increased, the rare cells 35 flow from the second outlet flow channel outlet 1521 into the first total outlet 19 of the upper cover plate 1 through the first cell outlet flow channel 26 of the flow guiding layer 2, the blood cells 36 on two sides flow into the flow channel outlet 16 through the first outlet flow channel 151 and the third outlet flow channel 153 respectively, and then flow into the second total outlet 20 through the second cell outlet flow channel 31, so that the rare cells 35 and the blood cells 36 are quickly and accurately sorted within a short distance, and due to the parallel arrangement of the sorting units 10, the flow channel flux is doubly improved, and the detection efficiency of the rare cells 35 is greatly improved.
In the present application, the viscoelastic sheath flow 34 and the viscoelastic sample flow 33 are both viscoelastic solutions, and the flow velocity of the viscoelastic sample flow 33 in the sample flow dual-split inlet flow channel 11 is consistent with the flow velocity of the viscoelastic sheath flow 34 in the sheath flow inlet flow channel 12. The speed of the cell with larger size is faster than that of the cell with smaller size which moves towards the center of the flow channel, so that rapid and accurate separation in a simple flow channel structure is realized, and the flow channels are arranged in parallel, so that the flow channel flux is improved in multiple, and the detection efficiency of rare cells is greatly improved.
The foregoing is only a preferred embodiment of the application, it being 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 present application, and such modifications and adaptations are intended to be comprehended within the scope of the application.
Claims (6)
1. A micro-fluidic chip for high-flux rapid and accurate cell sorting under low flow rate is characterized in that: including upper cover plate (1), guiding layer (2), runner layer (3) and lower apron (4) that set gradually from top to bottom, 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), wherein 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 inflow port (111); one end of the second cell outlet runner (31) is provided with a runner outlet (16), and the other end is provided with a runner layer cell outlet (32);
the sorting unit (10) comprises a sample flow double-split inlet runner (11), an extrusion straight runner (13), a shrinkage expansion array runner (14) and a sudden expansion sorting runner (15) which are connected in sequence;
the sample flow double-split inlet runner (11) comprises a first sample inflow runner (112), a sheath inflow runner (12) and a second sample inflow runner (113), wherein the inlet end of the first sample inflow runner (112) and the inlet end of the second sample inflow runner (113) are communicated with the sample inflow (111), and the outlet end of the first sample inflow runner (112), the outlet end of the second sample inflow runner (113) and the outlet end of the sheath inflow runner (12) are communicated with the inlet end of the extrusion straight runner (13); the inlet end of the sheath inflow channel (12) is provided with a sheath inflow (121), the sheath inflow channel (12) is positioned between the first sample inflow channel (112) and the second sample inflow channel (113), and the first sample inflow channel (112) and the second sample inflow channel (113) are symmetrical with respect to the sheath inflow channel (12);
the sudden expansion separation 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 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 all connected with the outlet end of the shrinkage 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 all connected with the 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 with respect to the second outlet flow channel (152);
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 guiding layer second outlet (28); one end of the sheath flow diversion layer inlet runner (23) is provided with a diversion layer sheath flow inlet (22), the other end is provided with a sheath flow branch inlet (24), one end of the first cell outlet runner (26) is provided with a runner branch outlet (25), and the other end is provided with a diversion layer first outlet (27);
the runner branch outlet (25) and the second outlet runner outlet (1521) are sequentially communicated from top to bottom;
the upper cover plate (1) is provided with a sample flow total inlet (17), a sheath flow total inlet (18), a first total outlet (19) and a second total outlet (20); the sample flow total inlet (17), the sample flow diversion layer inlet (21) and the sample flow channel layer inlet (29) are sequentially communicated from top to bottom; the sheath flow main inlet (18) and the diversion 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 total outlet (19) and the first outlet (27) of the diversion layer are sequentially communicated from top to bottom; the second total outlet (20), the second outlet (28) of the diversion layer and the cell outlet (32) of the runner layer are sequentially communicated from top to bottom;
the shrinkage-expansion array flow passage (14) is formed by alternately arranging and connecting a plurality of expansion flow passages and shrinkage flow passages, and the adjacent expansion flow passages and the shrinkage flow passages are mutually communicated.
2. The microfluidic chip for high-throughput rapid and accurate cell sorting at low flow rate according to claim 1, wherein the microfluidic chip is characterized in that: the sample inflow port (111), the sheath inflow port (121), the sheath inflow port runner (12), the extrusion straight runner (13), the shrinkage runner of the shrinkage expansion array runner (14), the sudden expansion sorting runner (15), the second outlet runner (152) and the runner outlet (16) are all on the same axis; the first sample inflow channel (112) is on the same side of the axis as the first outlet channel (151), and the second sample inflow channel (113) is on the other side of the axis as the third outlet channel (153).
3. The microfluidic chip for high-throughput rapid and accurate cell sorting at low flow rate according to claim 2, wherein the microfluidic chip is characterized in that: the shrinkage and expansion array flow passage (14) is of an axisymmetric structure.
4. The microfluidic chip for high-throughput rapid and accurate cell sorting at low flow rate according to claim 3, wherein the microfluidic chip is characterized in that: the included angle between the outlet section of the sheath inflow inlet flow channel (12) and the outlet section of the first sample inflow inlet flow channel (112) and the outlet section of the second sample inflow inlet flow channel (113) is an acute angle; the outlet section of the second outlet flow channel (152) forms an acute angle with the inlet section of the first outlet flow channel (151) and the inlet section of the third outlet flow channel (153), respectively.
5. The microfluidic chip for high-throughput rapid and accurate cell sorting at a low flow rate according to claim 4, wherein the microfluidic chip is characterized in that: the number of the sorting units (10) is 4, and the 4 sorting units (10) are arranged in parallel.
6. A high-throughput rapid and accurate cell sorting method under low flow rate is characterized in that: the microfluidic chip for high-throughput rapid and accurate cell sorting at low flow rate according to claim 1 comprises the following steps:
step 1, injecting a viscoelastic sample flow (33) with mixed blood from a sample flow total inlet (17) of an upper cover plate (1), flowing into a sample flow inlet (111) of a sample flow total inlet runner (30) of a runner 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 runner layer (3) through a sheath flow diversion layer inlet runner (23) of the diversion layer (2);
step 2, the viscoelastic sample flow (33) is extruded by elastic force directed to the wall surface before entering the shrinkage and expansion array flow channel (14) under the action of the viscoelastic sheath flow (34) and moves along the two side wall surfaces of the extrusion straight flow channel (13);
step 3, the cells enter the convergent-divergent array runner (14) to migrate towards the center of the runner under the action of the elastic force of viscoelastic fluid, the elastic force borne by large particles is larger, the cells migrate towards the center of the runner under the action of the drag force generated by the runner at the outlet of each divergent runner, the drag force borne by the cells is in direct proportion to the cell size, namely the drag force and the elastic force borne by rare cells (35) are larger, and the migration speed to the vicinity of the center of the runner is higher;
step 4: after flowing through the shrinkage and expansion array runner (14), rare cells (35) are positioned near the center of the runner and blood cells (36) are still positioned near the wall surface, after the cells enter the sudden expansion sorting runner (15), the distance between the rare cells (35) and the blood cells (36) is further enlarged in the sudden expansion sorting runner (15), the rare cells (35) flow into a first total outlet (19) of an upper cover plate (1) from a second outlet runner outlet (1521) through a first cell outlet runner (26) of a diversion layer (2), and the blood cells (36) at two sides flow into a second total outlet (20) of the upper cover plate (1) from a runner outlet (16) through a second cell outlet runner (31) respectively through a first outlet runner (151) and a third outlet runner (153), so that the rapid and accurate sorting of the rare cells (35) and the blood cells (36) in a short distance is realized.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101533230B1 (en) * | 2014-03-17 | 2015-07-02 | 연세대학교 산학협력단 | Multistage microfluidic chip and method for selective isolation of sample using the same |
| CN106190774A (en) * | 2016-07-22 | 2016-12-07 | 深圳睿思生命科技有限公司 | For capturing the micro-fluidic chip of circulating tumor cell and capture thereof and authentication method |
| CN111647499A (en) * | 2020-05-26 | 2020-09-11 | 东南大学 | Cell multistage sorting micro-fluidic chip integrating viscoelasticity and magnetic force |
| CN114073997A (en) * | 2021-11-30 | 2022-02-22 | 南京林业大学 | Microfluidic chip and method for realizing rapid and accurate cell sorting at low flow rate |
| WO2022088966A1 (en) * | 2020-10-29 | 2022-05-05 | 上海荧辉医疗器械有限公司 | Cell screening chip, and cell screening system and method |
-
2022
- 2022-12-09 CN CN202211577556.8A patent/CN115786074B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101533230B1 (en) * | 2014-03-17 | 2015-07-02 | 연세대학교 산학협력단 | Multistage microfluidic chip and method for selective isolation of sample using the same |
| CN106190774A (en) * | 2016-07-22 | 2016-12-07 | 深圳睿思生命科技有限公司 | For capturing the micro-fluidic chip of circulating tumor cell and capture thereof and authentication method |
| CN111647499A (en) * | 2020-05-26 | 2020-09-11 | 东南大学 | Cell multistage sorting micro-fluidic chip integrating viscoelasticity and magnetic force |
| WO2022088966A1 (en) * | 2020-10-29 | 2022-05-05 | 上海荧辉医疗器械有限公司 | Cell screening chip, and cell screening system and method |
| CN114073997A (en) * | 2021-11-30 | 2022-02-22 | 南京林业大学 | Microfluidic chip and method for realizing rapid and accurate cell sorting at low flow rate |
Non-Patent Citations (1)
| Title |
|---|
| Channel innovations for inertial microfluidics;Tang Wenlai;Lab Chip;第20卷(第19期);第3485-3502页 * |
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