Microfluidic sorting chip and system

Abstract

The utility model provides a microfluidic sorting chip and system, and relates to the technical field of particle sorting, the microfluidic sorting chip comprises a first sample inlet, a second sample inlet, a third sample inlet, a first sample outlet, a second sample outlet, a first flow channel and an annular flow channel; the first sample inlet is formed outside the annular flow channel, and the second sample inlet is formed inside the annular flow channel; two through-flow holes, namely a first through-flow hole and a second through-flow hole, are formed in the annular flow channel; the first through-flow hole is communicated with the first sample inlet, and the second through-flow hole is communicated with the second sample inlet; the first end of the first flow channel is communicated with the second through-flow hole; the second end of the first flow channel is communicated with the third sample inlet, the first sample outlet and the second sample outlet respectively. The micro-fluidic sorting chip and the micro-fluidic sorting system provided by the utility model have the advantages that the cell sorting operation is simple, the manufacturing process is simple, the target particles can be sorted from the cells to be sorted more accurately and efficiently, and the flexible sorting and nondestructive sorting of the cells can be realized.

Description

Technical Field

[0001] The utility model relates to the technical field of particle sorting, and in particular to a microfluidic sorting chip and system. Background Art

[0002] Microfluidic sorting chip is a device that can precisely manipulate fluids in micro-nanoscale space. Microfluidic sorting chip integrates basic operation units such as sample preparation, reaction, separation, detection, cell culture, sorting, lysis, etc. involved in the fields of chemistry and biology into a micrometer-scale chip, and uses the special properties of fluids in microchannels, such as laminar flow effect and diffusion effect, to manipulate and sort cells or particles.

[0003] In the process of using traditional microfluidic sorting chips for particle sorting in related technologies, there are defects such as complex operations. At the same time, traditional microfluidic sorting chips are dielectrophoresis sorting, and the electric field has an impact on the survival state of cells, and the survival rate of cells after sorting is not high. Therefore, how to reduce the complexity of particle sorting and achieve flexible sorting is a technical problem that technicians in this field need to solve urgently. Utility Model Content

[0004] The utility model provides a microfluidic sorting chip and system, which are used to solve the defects of complex operation and low cell survival rate in the process of particle sorting using traditional microfluidic sorting chips in the prior art, thereby reducing the complexity of particle sorting and realizing flexible sorting.

[0005] The utility model provides a microfluidic sorting chip, comprising: a first sample inlet, a second sample inlet, a third sample inlet, a first sample outlet, a second sample outlet, a first flow channel and an annular flow channel; the first sample inlet is arranged outside the annular flow channel, and the second sample inlet is arranged inside the annular flow channel; two through-holes are arranged on the annular flow channel, namely a first through-hole and a second through-hole; the first through-hole is connected to the first sample inlet, and the second through-hole is connected to the second sample inlet; the first end of the first flow channel is connected to the second through-hole, so that the sheath liquid flowing in from the first sample inlet is discharged into the second through-hole The hole covers the sample liquid flowing in from the second injection port, forming a mixed liquid of the sample liquid and the sheath liquid flowing into the first flow channel, and the sample liquid contains the particles to be sorted; the second end of the first flow channel is respectively connected to the third injection port, the first outlet and the second outlet, so that the mixed liquid is not impacted by the sorting liquid flowing in from the third injection port at the second end of the first flow channel, and flows into the waste liquid bottle through the second outlet, and the mixed liquid is impacted by the sorting liquid flowing in from the third injection port at the second end of the first flow channel, and flows into the liquid collection bottle through the first outlet.

[0006] According to a microfluidic sorting chip provided by the utility model, it also includes: a second flow channel, a third flow channel and a fourth flow channel; the second end of the first flow channel is connected to the third sample inlet through the second flow channel, the second end of the first flow channel is connected to the second sample outlet through the third flow channel, and the second end of the first flow channel is connected to the first sample outlet through the fourth flow channel.

[0007] According to a microfluidic sorting chip provided by the utility model, the first end of the fourth flow channel is connected to the second end of the first flow channel, the second end of the fourth flow channel is connected to the first sample outlet, and the diameter of the first end of the fourth flow channel is greater than the diameter of the second end of the fourth flow channel.

[0008] According to a microfluidic sorting chip provided by the utility model, it also includes: a fifth flow channel and a sixth flow channel; the first sample inlet is connected to the first flow hole through the fifth flow channel, and the second sample inlet is connected to the second flow hole through the sixth flow channel.

[0009] According to a microfluidic sorting chip provided by the utility model, the diameter of the flow channel in the microfluidic sorting chip is determined based on the diameter of the microparticles to be sorted.

[0010] According to a microfluidic sorting chip provided by the utility model, the material of the microfluidic sorting chip is polydimethylsiloxane.

[0011] According to a microfluidic sorting chip provided by the utility model, the particles to be sorted are cells, and the particles to be sorted exist in a cell matrix.

[0012] The utility model also provides a microfluidic sorting system, comprising: a microfluidic sorting chip as described above and a liquid driving device; the liquid driving device is used to drive a sample liquid containing particles to be sorted to flow from the second inlet of the microfluidic sorting chip into the microfluidic sorting chip, and drive a sheath liquid to flow from the first inlet of the microfluidic sorting chip into the microfluidic sorting chip, so that the sample liquid and the sheath liquid are mixed in the microfluidic sorting chip, and the liquid driving device is also used to drive a sorting liquid to flow from the third inlet of the microfluidic sorting chip into the microfluidic sorting chip, so that the mixed liquid of the sample liquid and the sheath liquid flows into a liquid collection bottle through the first outlet of the microfluidic sorting chip under the impact of the sorting liquid.

[0013] According to a microfluidic sorting system provided by the utility model, the liquid driving device includes: a pressurizing device; the pressurizing device is used to pressurize a first closed container containing the sheath liquid and a second closed container containing the sample liquid, respectively, so that the sheath liquid and the sample liquid flow into the first injection port and the second injection port respectively through a first pipe and a second pipe under the action of pressure, and the pressurizing device is also used to pressurize a third closed container containing the sorting liquid, so that the sorting liquid flows into the third injection port through a third pipe; the first pipe is used to connect the first closed container with the first injection port; the second pipe is used to connect the second closed container with the second injection port; the third pipe is used to connect the third closed container with the third injection port.

[0014] According to a microfluidic sorting system provided by the utility model, the pressurizing device includes a gas source and an air pump; the gas source is connected to the air pump, and the air pump is used to pressurize the first closed container to a first pressure value, the second closed container to a second pressure value, and the third closed container to a third pressure value by injecting different volumes of gas into the first closed container, the second closed container, and the third closed container respectively;

[0015] The first pressure value is calculated by the following formula:

[0016]

[0017] in, represents the first pressure value; represents the radius of the annular flow channel; represents the radius of the fifth flow channel; represents the specific resistance of the annular flow channel; represents the specific resistance of the fifth flow channel; represents the density of the sheath fluid; represents the expected flow rate of the sheath fluid in the fifth flow channel and the annular flow channel;

[0018] The second pressure value is calculated by the following formula:

[0019]

[0020] in, represents the second pressure value; represents the radius of the sixth flow channel; represents the specific resistance of the sixth flow channel; represents the density of the sample liquid; represents the expected flow rate of the sample liquid in the sixth flow channel;

[0021] The third pressure value is calculated by the following formula:

[0022]

[0023] in, represents the third pressure value; represents the radius of the second flow channel; represents the specific resistance of the second flow channel; represents the density of the separation liquid; represents the expected flow rate of the separation liquid in the second flow channel.

[0024] The microfluidic sorting chip and system provided by the utility model utilize an annular flow channel to achieve coating of the sample liquid by the sheath liquid, and respectively connect the annular flow channel and the third sample inlet, the first sample outlet and the second sample outlet through the two ends of the first flow channel, thereby achieving diversion of the mixed liquid of the sheath liquid and the sample liquid under the impact of the sorting liquid, and further achieving the sorting of the target particles. The microfluidic sorting chip has a simple structure, simple operation for cell sorting, and a simple manufacturing process, and can more accurately and efficiently sort the target particles from the cells to be sorted, can achieve flexible sorting and non-destructive sorting of cells, is particularly suitable for cell recovery and culture for further analysis, and has great application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] In order to more clearly illustrate the technical solutions in the present invention or the prior art, a brief introduction will be given below to the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0026] Figure 1 It is a structural schematic diagram of the microfluidic sorting chip provided by the utility model.

[0027] Figure 2 This is one of the schematic diagrams of simulation results of the microfluidic sorting chip provided by the utility model.

[0028] Figure 3 This is the second schematic diagram of the simulation results of the microfluidic sorting chip provided by the utility model.

[0029] Figure 4 It is a structural schematic diagram of the microfluidic sorting system provided by the utility model.

[0030] Figure 5 It is a working principle diagram of the liquid driving device in the microfluidic sorting system provided by the utility model.

[0031] Reference numerals:

[0032] 101: microfluidic sorting chip; 102: annular flow channel; 103: first flow hole; 104: second flow hole; 105: second injection port; 106: first injection port; 107: third injection port; 108: second outlet port; 109: first outlet port; 110: second flow channel; 111: third flow channel; 112: fourth flow channel; 113: fifth flow channel; 114: sixth flow channel; 115: first flow channel; 401: microfluidic sorting system; 402: liquid driving device. DETAILED DESCRIPTION

[0033] In order to make the purpose, technical solution and advantages of the utility model clearer, the technical solution of the utility model will be described clearly and completely in conjunction with the drawings in the utility model. Obviously, the described embodiments are part of the embodiments of the utility model, not all of the embodiments. Based on the embodiments in the utility model, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the utility model.

[0034] In the description of the utility model, it should be noted that, unless otherwise clearly specified and limited, the terms "installation", "connection" and "connection" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be indirectly connected through an intermediate medium, or it can be the internal communication of two components. For ordinary technicians in this field, the specific meanings of the above terms in this utility model can be understood according to specific circumstances.

[0035] In the description of the present application, the terms "first", "second", etc. are used to distinguish similar objects, and are not used to describe a specific order or sequence. It should be understood that the data used in this way can be interchangeable under appropriate circumstances, so that the embodiments of the present application can be implemented in an order other than those illustrated or described here, and the objects distinguished by "first", "second", etc. are usually a class, and the number of objects is not limited. For example, the first object can be one or more. In addition, in the description of the present application, "and / or" represents at least one of the connected objects, and the character " / " generally represents that the front and back associated objects are in an "or" relationship.

[0036] It should be noted that when traditional microfluidic sorting chips are used for particle sorting in related technologies, the connection methods of the microfluidic chips are diverse and the operation requirements are high. The detection process often requires multiple sampling steps, such as samples, quality control products / calibrators, diluents, cleaning solutions, and various reagents. This makes the operation of particle sorting using traditional microfluidic sorting chips more complicated.

[0037] In addition, traditional microfluidic sorting chips usually use steel needles to insert liquid pipes, without matching fixtures, and cannot be machined with threaded holes on the chip. The simple straight-in and straight-out method will lead to leakage and poor sealing. When the flow rate is too high, the interface is prone to leakage or even collapse.

[0038] In addition, traditional microfluidic sorting systems are usually suitable for oil phase droplets, that is, cells or viruses are encapsulated in droplets, and oil phase is used as the mobile phase. Most cells live in the cell matrix, which is more compatible with the human body environment. Some special cells (such as organoid cells) can only proliferate, differentiate and grow in three dimensions in the cell matrix (such as matrigel and collagen). However, the above special cells cannot be sorted in the oil phase.

[0039] Meanwhile, for large-sized particles such as organoid cells, the diameter of the large-sized particles is about 100-1000 However, traditional microfluidic sorting systems are difficult to sort large-sized particles.

[0040] Combine the following Figure 1-Figure 5 The present invention describes a microfluidic sorting chip.

[0041] Figure 1 This is a schematic diagram of the structure of the microfluidic sorting chip provided by the utility model. Figure 1 The microfluidic sorting chip provided by the utility model is described. Figure 1 As shown, the microfluidic sorting chip 101 includes: a first sample inlet 106, a second sample inlet 105, a third sample inlet 107, a first sample outlet 109, a second sample outlet 108, a first flow channel 115 and an annular flow channel 102; the first sample inlet 106 is arranged outside the annular flow channel 102, and the second sample inlet 105 is arranged inside the annular flow channel 102;

[0042] The annular flow channel 102 is provided with two through-holes, namely a first through-hole 103 and a second through-hole 104;

[0043] The first through hole 103 is connected to the first injection port 106, and the second through hole 104 is connected to the second injection port 105;

[0044] The first end of the first flow channel 115 is connected to the second flow hole 104, so that the sheath liquid flowing in from the first injection port 106 covers the sample liquid flowing in from the second injection port 105 at the second flow hole 104, forming a mixed liquid of the sample liquid and the sheath liquid flowing into the first flow channel 115, and the sample liquid contains the particles to be sorted;

[0045] The second end of the first flow channel 115 is connected to the third sample inlet 107, the first sample outlet 109 and the second sample outlet 108 respectively, so that the mixed liquid at the second end of the first flow channel 115 is not impacted by the sorting liquid flowing in from the third sample inlet 107, and flows into the waste liquid bottle through the second sample outlet 108, and the mixed liquid is impacted by the sorting liquid flowing in from the third sample inlet 107 at the second end of the first flow channel 115, and flows into the liquid collection bottle through the first sample outlet 109.

[0046] It should be noted that the particles to be sorted are the sorting objects of the microfluidic sorting system provided by the utility model. The particles to be sorted include the required target particles and non-target particles. The microfluidic sorting chip 101 provided by the utility model can sort the sample liquid containing the particles to be sorted into target liquid including target particles and waste liquid not including target particles, thereby realizing the sorting of the particles to be sorted.

[0047] It should be noted that the microparticles in the embodiments of the present invention may be cells, viruses, enzymes, proteins and other types of microparticles, and the cells may include single cells and organoid cells.

[0048] It can be understood that the particles to be sorted and the target particles in the embodiments of the present invention can be determined based on actual needs.

[0049] For example, the target particles in the embodiments of the present invention are organoid cells of a certain specific form, and the particles to be sorted include organoid cells of various forms. The microfluidic sorting system chip provided by the present invention can sort the sample liquid containing organoid cells of various forms into a target liquid including the organoid cells of the above-mentioned specific form and a waste liquid that does not include the organoid cells of the above-mentioned specific form.

[0050] For another example, the target particles in the embodiment of the present invention are a certain type of organoid cells, and the particles to be sorted include multiple types of organoid cells. The microfluidic sorting system chip provided by the present invention can sort the sample liquid containing multiple types of organoid cells into a target liquid including the above-mentioned specific type of organoid cells and a waste liquid not including the above-mentioned specific type of organoid cells. The particles to be sorted and the target particles are not specifically limited in the embodiment of the present invention.

[0051] The microfluidic sorting chip 101 integrates the basic operating units such as sample preparation, reaction, separation, detection, cell culture, sorting, lysis, etc. involved in the fields of chemistry and biology onto a micrometer-scale chip, and utilizes the special properties of the fluid in the microchannel, such as laminar flow effect and diffusion effect, to manipulate and sort cells or particles.

[0052] The microfluidic sorting chip 101 is mainly composed of a chip substrate, a fluid flow channel, a microstructure, an electrode layer, a dielectric layer, and a hydrophobic layer.

[0053] The chip substrate is the main structure of the microfluidic sorting chip 101, and is usually made of silicon, glass or polymer materials (such as PMMA, PDMS, etc.). These materials have good processing performance and chemical stability, and can withstand the pressure, temperature and chemical environment in microfluidic operations. Tiny fluid channels and reaction chambers are processed on the substrate to control and guide the flow of fluids and to sort cells or particles.

[0054] The fluid flow channel is the core part of the microfluidic sorting chip 101, which is used to transport and mix fluid samples. The size of these flow channels is usually at the micron level, which can accurately control the flow direction and speed of the fluid. The design of the flow channel can be customized according to the specific sorting requirements, such as setting a liquid reservoir, a main flow channel, a focusing flow channel, a detection flow channel and a collection flow channel, etc., to achieve accurate sorting and collection of cells.

[0055] The microstructures include micro-components such as reaction chambers, valves, pumps, and sensors. In the microfluidic sorting chip 101, these microstructures are used to control the flow and mixing of fluids, and to monitor the physical and chemical changes in the reaction process in real time. For example, valves and pumps can be used to control the flow speed and direction of the fluid to ensure uniform distribution and accurate sorting of cells in the flow channel. Sensors are used to monitor parameters such as cell concentration and flow rate in the fluid so as to adjust the sorting conditions in a timely manner.

[0056] As an optional embodiment, the material of the microfluidic sorting chip 101 is polydimethylsiloxane.

[0057] Specifically, the chip substrate of the microfluidic sorting chip 101 in the embodiment of the present invention is polydimethylsiloxane (PDMS).

[0058] The microfluidic sorting chip 101 in the embodiment of the present invention is equipped with three sample inlets and two sample outlets, namely a first sample inlet 106 , a second sample inlet 105 and a third sample inlet 107 , and a first sample outlet 109 and a second sample outlet 108 .

[0059] The first injection port 106 of the microfluidic sorting chip 101 is used to inject sheath liquid into the microfluidic sorting chip 101. The second injection port 105 of the microfluidic sorting chip 101 is used to inject sample liquid containing particles to be sorted into the microfluidic sorting chip 101. The third injection port 107 of the microfluidic sorting chip 101 is used to inject sorting liquid into the microfluidic sorting chip 101.

[0060] The first sample outlet 109 of the microfluidic sorting chip 101 is used for the outflow of the target liquid including the target particles, and the second sample outlet 108 of the microfluidic sorting chip 101 is used for the outflow of the waste liquid not including the target particles.

[0061] It should be noted that in the embodiment of the utility model, the sample liquid, sheath liquid and sorting liquid can be driven to flow into the microfluidic sorting chip 101 in a variety of ways. For example, in the embodiment of the utility model, a 2 / 50 mL centrifuge tube can be used to drive the sample liquid, sheath liquid and sorting liquid to flow into the microfluidic sorting chip 101. In the embodiment of the utility model, a pumping device (such as a syringe pump, a pressure pump, a pulsation-free pump) and other devices can also be used to drive the sample liquid, sheath liquid and sorting liquid to flow into the microfluidic sorting chip 101.

[0062] It should be noted that the timing of driving the sorting liquid to flow into the microfluidic sorting chip 101 in the embodiment of the utility model can be determined by a technician, can be determined by relying on traditional fluorescence activated cell sorting analysis, or can be determined by relying on deep learning technology. The timing of driving the sorting liquid to flow into the microfluidic sorting chip 101 in the embodiment of the utility model is not specifically limited.

[0063] Optionally, in an embodiment of the utility model, an image acquisition device can be used to capture an image of the mixed liquid in the first flow channel 115 at a preset frequency, and then image recognition can be performed on the image of the mixed liquid based on deep learning technology. When target particles are identified in the image of the mixed liquid, it is determined that the mixed liquid corresponding to the image of the mixed liquid includes target particles, and then the sorting liquid can be driven to flow from the third sampling port 107 of the microfluidic sorting chip 101 into the microfluidic sorting chip 101, so that the mixed liquid flows to the liquid collecting bottle through the first sampling port 109 of the microfluidic sorting chip 101 under the impact of the sorting liquid, thereby obtaining the target liquid including the target particles in the liquid collecting bottle.

[0064] It is understandable that when the sorting liquid is driven to flow into the microfluidic sorting chip 101 from the third sample inlet 107 of the microfluidic sorting chip 101, the sample liquid is not impacted by the sorting liquid, and the sample liquid will directly flow into the waste liquid bottle through the second sample outlet 108 of the microfluidic sorting chip 101.

[0065] Optionally, the annular flow channel 102 in the embodiment of the utility model may be a circular annular flow channel 102 , an elliptical or rectangular annular flow channel 102 , or a polygonal annular flow channel 102 . Figure 1 The annular flow channel 102 shown is a polygonal annular flow channel 102. The specific shape of the annular flow channel 102 is not limited in the embodiment of the utility model.

[0066] Preferably, the annular flow channel 102 in the embodiment of the present utility model is as follows Figure 1 The polygon shown is composed of a rectangle and a triangle.

[0067] As an optional embodiment, the microfluidic sorting chip 101 further includes: a second flow channel 110, a third flow channel 111 and a fourth flow channel 112;

[0068] The second end of the first flow channel 115 is connected to the third sample inlet 107 through the second flow channel 110 , the second end of the first flow channel 115 is connected to the second sample outlet 108 through the third flow channel 111 , and the second end of the first flow channel 115 is connected to the first sample outlet 109 through the fourth flow channel 112 .

[0069] As an optional embodiment, the first end of the fourth flow channel 112 is connected to the second end of the first flow channel 115 , the second end of the fourth flow channel 112 is connected to the first sample outlet 109 , and the diameter of the first end of the fourth flow channel 112 is greater than the diameter of the second end of the fourth flow channel 112 .

[0070] As an optional embodiment, the microfluidic sorting chip 101 further includes: a fifth flow channel 113 and a sixth flow channel 114;

[0071] The first injection port 106 is connected to the first through-hole 103 through the fifth flow channel 113 , and the second injection port 105 is connected to the second through-hole 104 through the sixth flow channel 114 .

[0072] Optionally, the cross-sectional shape of the flow channel in the microfluidic sorting chip 101 in the embodiment of the utility model can be circular, rectangular, trapezoidal, semicircular, etc. The cross-sectional shape of the flow channel in the microfluidic sorting chip 101 is not limited in the embodiment of the utility model.

[0073] It can be understood that the flow channels in the microfluidic sorting chip 101 in the embodiment of the present invention include a first flow channel 115 , a second flow channel 110 , a third flow channel 111 , a fourth flow channel 112 , a fifth flow channel 113 and a sixth flow channel 114 .

[0074] As an optional embodiment, the diameter of the flow channel in the microfluidic sorting chip 101 is determined based on the diameter of the microparticles to be sorted.

[0075] It is understandable that the diameter of the flow channel in the microfluidic sorting chip 101 is larger than the diameter of the particle to be sorted. For example, the diameter of the particle to be sorted is 180 In the case of, the diameter of the flow channel in the microfluidic sorting chip 101 can be 200 .

[0076] It should be noted that the flow channel in the microfluidic sorting chip 101 in the embodiment of the present invention can be designed differently to achieve the separation of particles of different sizes (100-1000 μm). For example, when the length of the microfluidic sorting chip 101 and the depth of the flow channel are both 30 In the case of The length of the microfluidic sorting chip 101 and the depth of the flow channel are 200 In the case of That is, by changing the size of the flow channel in the microfluidic sorting chip 101, the sorting of particles of different sizes can be achieved, which can be applicable to various sizes, and is particularly suitable for the sorting of large-sized particles (such as organoid cells).

[0077] Optionally, the diameter of the first end of the fourth flow channel 112 may be determined based on the diameter of the second end of the fourth flow channel 112. For example, the diameter of the first end of the fourth flow channel 112 may be twice the diameter of the second end of the fourth flow channel 112. In the case of .

[0078] Alternatively, if Figure 1 As shown, the first end of the fourth flow channel 112 is funnel-shaped, and the diameter of the first end of the fourth flow channel 112 gradually decreases to the same diameter as the second end of the fourth flow channel 112 as it moves away from the second end of the first flow channel 115. The length of the first end of the fourth flow channel 112 can be determined based on prior knowledge and / or actual conditions, and the length of the first end of the fourth flow channel 112 is not limited in the embodiment of the utility model.

[0079] The design that the diameter of the first end of the fourth flow channel 112 is larger than the diameter of the second end helps to gradually focus and accelerate the fluid during the sorting process, thereby improving the accuracy and efficiency of sorting. At the same time, it can also reduce the diffusion and loss of the fluid in the flow channel, further improving the utilization rate of the reagent.

[0080] Alternatively, if Figure 1 As shown, the first flow channel 115 , the second flow channel 110 , the third flow channel 111 , the fifth flow channel 113 and the sixth flow channel 114 are all straight flow channels, and the fourth flow channel 112 is in a corner shape.

[0081] Alternatively, if Figure 1 As shown, the first flow channel 115 is located on the central axis of the annular flow channel 102 , the fifth flow channel 113 , the sixth flow channel 114 , the first flow channel 115 and the third flow channel 111 are coaxially arranged, and the second flow channel 110 and the first end of the fourth flow channel 112 are coaxially arranged.

[0082] The sheath liquid flows into the annular flow channel 102 from the first injection port 106 and the fifth flow channel 113. The sample liquid continues to flow in the annular flow channel 102. When it flows to the second through-hole 104, it can cover the sample liquid that flows to the second through-hole 104 from the second injection port 105 and the sixth flow channel 114. After the mixed liquid of the sample liquid and the sheath liquid flows in the first flow channel 115 to the second end of the first flow channel 115, it can flow to the liquid collection bottle through the fourth flow channel 112 and the first sample outlet 109 under the impact of the sorting liquid that flows in through the third injection port 107 and the second flow channel 110. When the mixed liquid of the sample liquid and the sheath liquid is not impacted by the sorting liquid, it can flow to the waste liquid bottle through the third flow channel 111 and the second sample outlet 108.

[0083] The microfluidic sorting chip in the embodiment of the utility model utilizes the annular flow channel 102 to achieve the coating of the sample liquid by the sheath liquid. The annular flow channel 102 and the third sample inlet, the first sample outlet and the second sample outlet are respectively connected through the two ends of the first flow channel, so that the mixed liquid of the sheath liquid and the sample liquid is diverted under the impact of the sorting liquid, thereby achieving the sorting of the target particles. The microfluidic sorting chip has a simple structure, simple operation for cell sorting, and a simple manufacturing process. It can more accurately and efficiently sort the target particles from the cells to be sorted, can achieve flexible sorting and non-destructive sorting of cells, is particularly suitable for cell recovery and culture for further analysis, and has great application prospects.

[0084] The microfluidic sorting chip in the embodiment of the utility model highly integrates multiple components such as sample inlets, sample outlets, flow channels and flow holes on a tiny chip, realizing highly integrated sample processing. This design not only reduces the size of the chip, but also improves the portability and operability of the chip. The microfluidic sorting chip modularizes the functions of coating, mixing, sorting and collecting sample liquids through different flow channels and connection structures, so that each functional unit can work independently without interfering with each other. This design improves the flexibility and versatility of the chip, making it convenient for users to select and adjust according to experimental needs.

[0085] The sheath liquid flowing into the microfluidic sorting chip in the embodiment of the utility model through the first inlet covers the sample liquid flowing in from the second inlet at the second flow hole to form a mixed liquid, which helps to reduce the diffusion and loss of the sample liquid in the flow channel and improve the accuracy of sorting. At the same time, the sheath liquid can also protect and stabilize the sample liquid to prevent it from being damaged during the sorting process. The mixed liquid is impacted by the sorting liquid flowing in from the third inlet at the second end of the first flow channel, thereby realizing the sorting of the particles to be sorted. This impact method is efficient and fast, and can significantly improve the sorting efficiency. At the same time, by adjusting the flow rate and impact strength of the sorting liquid, it is possible to achieve accurate sorting of particles of different sizes and properties.

[0086] The flow channel design in the microfluidic sorting chip in the embodiment of the utility model has been carefully optimized, including the size, curvature and connection structure of the flow channel, to ensure smooth flow and uniform distribution of the fluid in the flow channel, which helps to reduce the resistance and loss of the fluid in the flow channel and improve the utilization and sorting efficiency of the fluid.

[0087] Due to the integrated and miniaturized design of the microfluidic sorting chip in the embodiment of the utility model, the required amount of reagents is also greatly reduced, which helps to reduce the experimental cost and reduce the pollution to the environment. In addition, by accurately controlling the amount and flow rate of the reagents, the accuracy and reliability of the experiment can be further improved.

[0088] The microfluidic sorting chip in the embodiment of the utility model is small in size, easy to carry and store, and can be used in different experimental environments, which improves the flexibility and convenience of the experiment. Due to the highly integrated and automated design of the microfluidic sorting chip, the experimental operation becomes simple and easy to understand. The user only needs to add samples and reagents according to the specified steps to automatically complete the sorting operation. This design reduces the requirements for the professionalism and experience accumulation of the operator, allowing more people to use this chip for experiments and research.

[0089] As an optional embodiment, when the particles to be sorted are cells, the cells to be sorted live in a cell matrix.

[0090] The particles to be sorted in the embodiment of the utility model exist in the cell matrix, which can realize the aqueous phase sorting of the particles to be sorted, can be better applicable to the needs that cannot be sorted in the oil phase, and can be applicable to the sorting of various biological components (such as cells, viruses, enzymes, proteins, etc.) in the aqueous phase at the same time.

[0091] By performing physical simulation on the structure of the microfluidic sorting chip 101 in the microfluidic sorting system, it can be verified that the microfluidic sorting chip 101 has no structural defects and other potential design defects in fluid mechanics, and the design of the microfluidic sorting chip 101 can be optimized to ensure the stability and reliability of the microfluidic sorting system in practical applications. Figure 2 This is one of the schematic diagrams of simulation results of the microfluidic sorting chip provided by the utility model. Figure 3 The second schematic diagram of the simulation results of the microfluidic sorting chip provided by the utility model is shown in FIG. Figure 2 and Figure 3 shown.

[0092] Figure 4 This is a schematic diagram of the structure of the microfluidic sorting system provided by the utility model. Figure 4 The microfluidic sorting system provided by the utility model is described. Figure 4As shown, a microfluidic sorting system 401 comprises: any of the above microfluidic sorting chips 101 and a liquid driving device 402;

[0093] The liquid driving device 402 is used to drive the sample liquid containing the particles to be sorted to flow from the second inlet 105 of the microfluidic sorting chip 101 into the microfluidic sorting chip 101, and drive the sheath liquid to flow from the first inlet 106 of the microfluidic sorting chip 101 into the microfluidic sorting chip 101, so that the sample liquid and the sheath liquid are mixed in the microfluidic sorting chip 101. The liquid driving device 402 is also used to drive the sorting liquid to flow from the third inlet 107 of the microfluidic sorting chip 101 into the microfluidic sorting chip 101, so that the mixed liquid of the sample liquid and the sheath liquid flows into the liquid collection bottle through the first outlet 109 of the microfluidic sorting chip 101 under the impact of the sorting liquid.

[0094] As an optional embodiment, the liquid driving device 402 includes: a pressurizing device;

[0095] The pressurizing device is used to pressurize the first sealed container containing the sheath liquid and the second sealed container containing the sample liquid, respectively, so that the sheath liquid and the sample liquid flow into the first injection port 106 and the second injection port 105 through the first pipe and the second pipe under the action of pressure, respectively. The pressurizing device is also used to pressurize the third sealed container containing the sorting liquid, so that the sorting liquid flows into the third injection port 107 through the third pipe;

[0096] The first pipeline is used to connect the first sealed container with the first injection port 106 ; the second pipeline is used to connect the second sealed container with the second injection port 105 ; the third pipeline is used to connect the third sealed container with the third injection port 107 .

[0097] As an optional embodiment, the pressurizing device includes an air source and an air pump; the air source is connected to the air pump, and the air pump is used to inject different volumes of gas into the first closed container, the second closed container and the third closed container respectively, so that the first closed container is pressurized to a first pressure value, the second closed container is pressurized to a second pressure value, and the third closed container is pressurized to a third pressure value.

[0098] Figure 5 This is a working principle diagram of the liquid driving device in the microfluidic sorting system provided by the utility model. Figure 5 As shown, the working principle of the liquid driving device 402 in the embodiment of the utility model is: a certain amount of solution is contained in a closed container, and a gas with a certain pressure is introduced into the closed container. The solution in the closed container will enter the microfluidic sorting chip 101 along the liquid pipeline under the action of pressure.

[0099] Optionally, the pressurizing device in the embodiment of the utility model may include an air source and an air pump, the air source is connected to the air pump, and the air pump increases the pressure in the closed container by injecting gas in the air source into the closed container.

[0100] Optionally, the gas source may be nitrogen or an air compressor. The air pump may be a Fluigent air pump.

[0101] The air pump can pressurize the first closed container to a first pressure value, the second closed container to a second pressure value, and the third closed container to a third pressure value by injecting different volumes of gas into the first closed container, the second closed container and the third closed container respectively.

[0102] Optionally, precision pressure sensors are provided at the first injection port 106, the second injection port 105 and the third injection port 107 to monitor the pressure at the first injection port 106, the second injection port 105 and the third injection port 107, so as to adjust the volume of gas injected into the first closed container, the second closed container and the third closed container by the air pump.

[0103] It should be noted that the pressurizing device in the embodiment of the utility model can generate different air pressures.

[0104] Accordingly, the first pressure value that the pressurizing device pressurizes into the first closed container can be determined based on the diameter and specific resistance of the fifth flow channel 113 and the annular flow channel 102 , the density of the sheath fluid, and the expected flow rate of the sheath fluid in the fifth flow channel 113 and the annular flow channel 102 .

[0105] The second pressure value that the pressurizing device applies pressure to the second closed container may be determined based on the diameter and specific resistance of the sixth flow channel 114 , the density of the sample liquid, and the expected flow rate of the sample liquid in the sixth flow channel 114 .

[0106] The third pressure value applied by the pressurizing device to the third closed container may be determined based on the diameter and specific resistance of the second flow channel 110 , the density of the separation liquid, and the expected flow rate of the separation liquid in the second flow channel 110 .

[0107] The first pressure value is calculated by the following formula:

[0108]

[0109] in, Indicates the first pressure value; represents the radius of the annular flow channel 102; represents the radius of the fifth flow channel 113; represents the specific resistance of the annular flow channel 102; represents the specific resistance of the fifth flow channel 113; represents the density of the sheath fluid; denoted the expected flow rate of the sheath fluid in the fifth flow channel 113 and the annular flow channel 102 .

[0110] The second pressure value is calculated by the following formula:

[0111]

[0112] in, Indicates the second pressure value; represents the radius of the sixth flow channel 114; represents the specific resistance of the sixth flow channel 114; Indicates the density of the sample liquid; represents the expected flow rate of the sample liquid in the sixth flow channel 114 .

[0113] The third pressure value is calculated by the following formula:

[0114]

[0115] in, Indicates the third pressure value; represents the radius of the second flow channel 110; represents the specific resistance of the second flow channel 110; Indicates the density of the sorting liquid; represents the expected flow rate of the separation liquid in the second flow channel 110 .

[0116] It should be noted that the expected flow rates of the sheath fluid in the fifth flow channel 113 and the annular flow channel 102 , the expected flow rate of the sample fluid in the sixth flow channel 114 , and the expected flow rate of the sorting fluid in the second flow channel 110 may be determined based on prior knowledge and / or actual conditions.

[0117] Optionally, a solenoid valve may be provided between the air pump and the sealed container, and the volume of gas input into the sealed container by the air pump may be controlled by the solenoid valve.

[0118] The liquid driving device in the embodiment of the utility model can accurately pressurize the closed container containing sheath liquid, sample liquid and sorting liquid, ensure that the liquid flows stably into the corresponding sampling port under the action of pressure, and realize high-precision control of the liquid flow rate in the microfluidic sorting chip. By accurately controlling the flow rate of the sheath liquid, sample liquid and sorting liquid in the microfluidic sorting chip, the analysis throughput and efficiency of the microfluidic sorting chip can be improved. The use of closed containers and piping systems can effectively prevent the contamination and volatilization of the sheath liquid, sample liquid and sorting liquid during the flow process, and ensure the purity and stability of the sheath liquid, sample liquid and sorting liquid. The liquid driving device has a relatively simple structure and is relatively easy to maintain and maintain, which reduces the operating cost of the microfluidic sorting system and improves the reliability and service life of the microfluidic sorting system.

[0119] It should be noted that in the microfluidic sorting system 401 provided by the present invention, below the bright field light source is the microfluidic sorting chip 101, and the flow of the sample liquid, sheath liquid and sorting liquid in the microfluidic sorting chip 101 is controlled by the liquid driving device 402. Below the microfluidic sorting chip 101 is an optical module, which is composed of an objective lens and a filter, and the fluorescent field light source is located in the middle of the image acquisition device. The microfluidic sorting system 401 has the advantages of simple operation, fast detection and high precision.

[0120] The performance of the microfluidic sorting system 401 provided by the utility model is verified through sorting experiments, including sorting efficiency, accuracy, repeatability, etc. According to the experimental feedback, the design and hardware configuration of the microfluidic sorting system 401 are continuously adjusted and optimized until the microfluidic sorting system 401 reaches the expected performance indicators.

[0121] The microfluidic sorting system 401 provided by the present invention can determine the diameter of the flow channel in the microfluidic sorting chip 101 based on the diameter of the particles to be sorted, and is suitable for sorting particles of different sizes to be sorted, for example, it can realize the sorting of single cells and organoid cells respectively.

[0122] The microfluidic sorting system 401 provided by the utility model uses an air pump pressurization method to control the flow of sheath liquid, sample liquid and sorting liquid in the microfluidic sorting chip 101, which can realize water phase sorting and is suitable for sorting various biological components (such as cells, viruses, enzymes, proteins, etc.) in the water phase at the same time.

[0123] The microfluidic sorting system 401 provided by the present invention can realize flexible sorting without loss, and is particularly suitable for cell recovery and culture for further analysis.

[0124] In the microfluidic sorting system provided by the embodiment of the utility model, the microfluidic sorting chip and the liquid driving device are closely combined to form a complete sorting system. This highly integrated design makes the system more compact, lightweight, easy to carry and operate. The microfluidic sorting system realizes the automatic inflow and mixing of sample liquid, sheath liquid and sorting liquid through the pressurization of the liquid driving device. The user only needs to set the relevant parameters, and the microfluidic sorting system can automatically complete the sorting process, greatly reducing the difficulty of operation and labor intensity. The microfluidic sorting system can adjust the flow rate, pressure and other parameters of the sample liquid, sheath liquid and sorting liquid according to different experimental requirements to adapt to different sorting conditions and particle characteristics. The design of the microfluidic sorting chip is easy to expand and modify, and more sample inlets, sample outlets and flow channels can be added to meet more complex sorting requirements. At the same time, the liquid driving device can also be upgraded and expanded as needed to improve the performance and function of the system. The structural design of the microfluidic sorting system is reasonable and easy to disassemble and clean. This helps to extend the service life of the system and keep it in good working condition. The liquid drive device adopts mature technical means such as pressurizing device and air pump, which has stable and reliable performance. The microfluidic sorting system can also be equipped with corresponding safety protection measures, such as pressure sensor, overcurrent protection, etc., to ensure the safe operation of the system.

[0125] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the utility model, rather than to limit it. Although the utility model has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the various embodiments of the utility model.

Claims

1. A microfluidic sorting chip, characterized in that: include: A first injection port, a second injection port, a third injection port, a first outlet port, a second outlet port, a first flow channel and an annular flow channel; the first injection port is arranged outside the annular flow channel, and the second injection port is arranged inside the annular flow channel; The annular flow channel is provided with two through-flow holes, namely a first through-flow hole and a second through-flow hole; The first flow hole is connected to the first injection port, and the second flow hole is connected to the second injection port; The first end of the first flow channel is connected to the second flow hole, so that the sheath liquid flowing in from the first injection port covers the sample liquid flowing in from the second injection port at the second flow hole, forming a mixed liquid of the sample liquid and the sheath liquid flowing into the first flow channel, and the sample liquid contains the particles to be sorted; The second end of the first flow channel is connected to the third sample inlet, the first sample outlet and the second sample outlet respectively, so that the mixed liquid flows into the waste liquid bottle through the second sample outlet without being impacted by the sorting liquid flowing in from the third sample inlet at the second end of the first flow channel, and the mixed liquid flows into the liquid collection bottle through the first sample outlet when being impacted by the sorting liquid flowing in from the third sample inlet at the second end of the first flow channel.

2. The microfluidic sorting chip according to claim 1, characterized in that: Also includes: a second flow channel, a third flow channel, and a fourth flow channel; The second end of the first flow channel is connected to the third sample inlet through the second flow channel, the second end of the first flow channel is connected to the second sample outlet through the third flow channel, and the second end of the first flow channel is connected to the first sample outlet through the fourth flow channel.

3. The microfluidic sorting chip according to claim 2, characterized in that: The first end of the fourth flow channel is connected to the second end of the first flow channel, the second end of the fourth flow channel is connected to the first sample outlet, and the diameter of the first end of the fourth flow channel is greater than the diameter of the second end of the fourth flow channel.

4. The microfluidic sorting chip according to claim 3, characterized in that: Also includes: a fifth flow channel and a sixth flow channel; The first injection port is connected to the first flow hole through the fifth flow channel, and the second injection port is connected to the second flow hole through the sixth flow channel.

5. The microfluidic sorting chip according to claim 4, characterized in that: The diameter of the flow channel in the microfluidic sorting chip is determined based on the diameter of the microparticles to be sorted.

6. The microfluidic sorting chip according to claim 4, characterized in that: The material of the microfluidic sorting chip is polydimethylsiloxane.

7. The microfluidic sorting chip according to any one of claims 4 to 6, characterized in that: The particles to be sorted are cells, and the particles to be sorted exist in a cell matrix.

8. A microfluidic sorting system, characterized in that: include: The microfluidic sorting chip and liquid driving device according to any one of claims 4 to 7; The liquid driving device is used to drive the sample liquid containing the particles to be sorted to flow into the microfluidic sorting chip from the second injection port of the microfluidic sorting chip, and drive the sheath liquid to flow into the microfluidic sorting chip from the first injection port of the microfluidic sorting chip, so that the sample liquid and the sheath liquid are mixed in the microfluidic sorting chip. The liquid driving device is also used to drive the sorting liquid to flow into the microfluidic sorting chip from the third sampling port of the microfluidic sorting chip, so that the mixed liquid of the sample liquid and the sheath liquid flows into the liquid collection bottle through the first sampling port of the microfluidic sorting chip under the impact of the sorting liquid.

9. The microfluidic sorting system according to claim 8, characterized in that: The liquid driving device comprises: a pressurizing device; The pressurizing device is used to pressurize the first sealed container containing the sheath liquid and the second sealed container containing the sample liquid, respectively, so that the sheath liquid and the sample liquid flow into the first injection port and the second injection port respectively through the first pipe and the second pipe under the action of pressure. The pressurizing device is also used to pressurize the third closed container containing the separation liquid, so that the separation liquid flows into the third injection port through the third pipeline; The first pipeline is used to connect the first sealed container with the first injection port; the second pipeline is used to connect the second sealed container with the second injection port; the third pipeline is used to connect the third sealed container with the third injection port.

10. The microfluidic sorting system according to claim 9, characterized in that: The pressurizing device comprises an air source and an air pump; the air source is connected to the air pump, and the air pump is used to pressurize the first sealed container to a first pressure value, the second sealed container to a second pressure value, and the third sealed container to a third pressure value by injecting different volumes of gas into the first sealed container, the second sealed container, and the third sealed container, respectively; The first pressure value is calculated by the following formula: ； in, represents the first pressure value; represents the radius of the annular flow channel; represents the radius of the fifth flow channel; represents the specific resistance of the annular flow channel; represents the specific resistance of the fifth flow channel; represents the density of the sheath fluid; represents the expected flow rate of the sheath fluid in the fifth flow channel and the annular flow channel; The second pressure value is calculated by the following formula: ； in, represents the second pressure value; represents the radius of the sixth flow channel; represents the specific resistance of the sixth flow channel; represents the density of the sample liquid; represents the expected flow rate of the sample liquid in the sixth flow channel; The third pressure value is calculated by the following formula: ； in, represents the third pressure value; represents the radius of the second flow channel; represents the specific resistance of the second flow channel; represents the density of the separation liquid; represents the expected flow rate of the separation liquid in the second flow channel.

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