Micro-fluidic chip for skin cell sorting

Abstract

The utility model provides a microfluidic chip for skin cell sorting, which relates to the technical field of cell sorting and comprises a substrate, a microfluidic channel is arranged on the upper surface of the substrate, and the microfluidic channel comprises a primary sorting channel and two secondary sorting channels. The first-stage sorting channel is composed of three sorting matrixes, the second-stage sorting channel is composed of two sorting matrixes, and each sorting matrix is formed by connecting an expansion area with a contraction area. According to the skin cell sorting device, an external magnetic field acts on the immunomagnetic beads and combined cells, so that the sorting efficiency is greatly improved under the condition of effectively sorting the skin cells. Moreover, corollary equipment is simplified and only relates to the upright microscope and the micro-injection pump, so that the operation of the micro-fluidic chip is simpler and more convenient, and the use cost is remarkably reduced.

Description

Technical Field

[0001] This utility model relates to the field of cell sorting technology, and more specifically, to a microfluidic chip for skin cell sorting. Background Technology

[0002] Microfluidic chips utilize microelectromechanical fabrication (MEMS) technology to construct complex microchannels on a chip, enabling precise manipulation of minute amounts of fluids. Since the vast majority of cells are on the micrometer scale, their size is well-suited to the microchannel dimensions, allowing for the manipulation of even a few or single cells. Furthermore, the small volume of the microchannels helps shorten mass transfer time, accelerate time response, and maintain long-term dynamic tracking. Microfluidic chips are widely used in cell mixing, focusing, sorting, and detection, covering fields such as separation and analysis, chemical synthesis, medical diagnostics, cell biology, neurobiology, systems biology, structural biology, and microbiology. Miniaturized, integrated, and high-throughput microfluidic chips, known as lab-on-a-chip, have broad application prospects in life science research and clinical diagnostics.

[0003] In the field of skin tissue engineering, the isolation and enrichment of target cells are prerequisites for targeted research. Existing skin cell sorting technologies, such as flow cytometry, immunomagnetic bead sorting, and differential adhesion, while effective in specific applications, face a series of challenges. For example, flow cytometry is complex, requires a large number of cell samples, and relies on costly equipment and specialized operation. While immunomagnetic bead sorting is highly efficient in cell enrichment, its surface functionalization process is cumbersome and susceptible to external interference, and it lacks selectivity for certain cell types. Differential adhesion, although capable of physical cell separation, requires long-term culture and is complex to operate. These limitations restrict the ability to efficiently isolate target cells from rare cell sources or limited samples. To address the shortcomings of existing technologies, this invention provides a novel microfluidic chip combining microfluidic sorting and immunomagnetic bead technology, aiming to solve the challenges of skin cell sorting. This technology achieves precise manipulation of microfluidics by constructing complex microchannels on the chip, thereby accurately and efficiently separating and enriching target cells. This method not only significantly reduces the required cell quantity, but also offers fast analysis speed, simple operation, and low cost. Utility Model Content

[0004] This invention provides a microfluidic chip for skin cell sorting, aiming to overcome the shortcomings of traditional skin cell sorting methods, simplify the sorting process, and improve sorting efficiency and accuracy.

[0005] To achieve the above objectives, the present invention adopts the following technical solution:

[0006] A microfluidic chip for skin cell sorting, the microfluidic chip is made of a glass substrate, on which microfluidic channels are formed, and the microfluidic channels are provided with a width of 30-60um.

[0007] Preferably, the microfluidic channel includes one primary sorting channel and two secondary sorting channels. Preferably, the primary sorting channel consists of three linearly arranged sorting matrices, while the secondary sorting channels consist of two linearly arranged sorting matrices.

[0008] Preferably, each sorting matrix consists of an extended portion connected to a contracted portion, the contracted portion having a width of 30-60 μm and a length of 120-200 μm, and the extended portion having a width of 250-500 μm and a length of 600-800 μm.

[0009] Preferably, the microfluidic channel also includes a sample inlet channel connected to the primary sorting channel, wherein the width of the sample inlet channel is 300-500 μm.

[0010] Preferably, the microfluidic channel also includes a rinsing channel connected to the primary sorting channel via rinsing holes, wherein the width of the rinsing channel is 300-500 μm.

[0011] Preferably, the angle between the sample inlet channel and the rinsing channel is 70-90°.

[0012] In addition, the microfluidic channel includes two flow channels connected to the primary sorting channel and the secondary sorting channel, with the angle between the two flow channels being 90-120°.

[0013] The first and second-level sorting channels are connected to two first collection holes through two collection channels, wherein the included angle between the two collection channels is 60-80°.

[0014] The second secondary sorting channel connects two second collection holes through two collection channels, wherein the included angle between the two collection channels is 60-80°.

[0015] Because of the above-mentioned technical solution, the microfluidic chip of this invention has a reasonable design structure and is easy to operate, and can effectively achieve efficient sorting of skin cells, providing new tools and methods for research and clinical diagnosis in related fields. Attached Figure Description

[0016] To more clearly illustrate the technical solutions in the embodiments of this utility model, the drawings used in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this utility model. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0017] Figure 1 This is a two-dimensional structural schematic diagram of a microfluidic chip for skin cell sorting according to an embodiment of the present invention;

[0018] Explanation of reference numerals in the attached figures:

[0019] 1-Substrate; 11-Flow channel layer; 12-Substrate layer; 21-Primary sorting channel; 22, 23-Secondary sorting channels; 24-Flow channel; 25-Rinsing channel; 26, 27-Flow channels; 4-Sample hole; 5-Rinsing hole; 61, 62, 63, 64-Sample collection hole; 71-Mobile phone channel.

[0020] Figure 2 This is a schematic diagram of the primary sorting channel structure of the microfluidic chip for skin cell sorting according to an embodiment of the present invention;

[0021] Explanation of reference numerals in the attached figures:

[0022] 3-Sorting matrix; 31-Extended part; 32-Contracted part. Detailed Implementation

[0023] To make the above-mentioned objectives, features, and advantages of this utility model more apparent and understandable, the technical solutions in specific embodiments of this utility model will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only a part of the embodiments of this utility model, and not all of them. All other embodiments obtained by those skilled in the art based on the embodiments of this utility model without creative effort are within the scope of protection of this utility model.

[0024] It should be noted that the technical terms used in the specification and claims of this utility model should have the ordinary meaning understood by a person skilled in the art to which this utility model pertains. The terms "comprising" or "including," and similar words used in the specification and claims, mean that the components or objects preceding "comprising" or "including" cover the components or objects listed after "comprising" or "including" and their equivalents, and do not exclude other components or objects. Terms such as "connected" or "linked" are not limited to physical or mechanical connections, nor are they limited to direct or indirect connections.

[0025] It should be understood that the descriptions of orientation in the specification, claims, and drawings of this utility model, such as front, back, up, down, etc., indicating orientation or positional relationships, are based on the orientation or positional relationships shown in the drawings and are only for the convenience of describing this utility model and simplifying the description. They are not intended to indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation on this utility model.

[0026] This invention provides a microfluidic chip for skin cell sorting, see [link to relevant documentation]. Figure 1 It includes a base 1, on which a section with a height of 50 is formed. The microfluidic channel, comprising an injection channel 24, a rinsing channel 25, a primary sorting channel 21, two flow channels 26 and 27, two secondary sorting channels 22 and 23, and a collection channel 71, is sequentially connected along the flow direction of the cell suspension (as referred to in the context of this invention). Specifically, the outlet of the injection channel 24 is connected to the inlet of the primary sorting channel 21, and the outlet of the primary sorting channel 21 is connected to the inlet of the flow channels 26 and 27. The primary sorting channel 21 is used to gradually separate the cell suspension into single-cell droplets, achieving orderly arrangement of cells in the primary sorting channel 21, especially in the latter half, thereby sorting out the cells to be selected. The collection channel 71 is used to collect the separated cells to be selected, thus forming a fluid-flowing microfluidic channel and constituting a cell collection area on the substrate 1.

[0027] like Figure 1 As shown, the sample inlet channel 24 has a width of 400 μm, the rinsing channel 25 has a width of 400 μm, and the angle between the sample inlet channel 24 and the rinsing channel 25 is 80°. The primary sorting channel 21 consists of three sorting matrices 3 arranged linearly, and the secondary sorting channels 22 and 23 consist of two sorting matrices 3 arranged linearly. Each sorting matrix 3 consists of an extension portion 31 connected to a contraction portion 32, where the contraction portion 32 has a width of 50 μm and a length of 150 μm, and the extension portion 31 has a width of 350 μm and a length of 700 μm. The angle between the two flow channels 26 and 27 is 100°. Two collection channels connect to two collection holes 61 and 62, where the included angle between the two collection channels is 70°. The primary and secondary sorting channels 22 and 23 are connected to the two collection holes 61 and 62 via the two collection channels, where the included angle between the two collection channels is 70°.

[0028] In practical use, sample well 4 is connected to a syringe via a conduit, and the outlet of collection channel 71 is connected to a cell culture device such as a test tube, culture dish, or cell culture flask via a conduit. It is important to note that each inlet and outlet on the microfluidic chip is connected to its associated microfluidic channel, and simultaneously, each inlet and outlet is connected to an external fluid-driven sample injection device such as a micro-injection pump via a conduit to provide the driving force for fluid flow. Before sample injection, the microfluidic chip of this invention is placed on the stage of an upright microscope such as a fluorescence microscope, with the intersection of sorting channel 21 and flow channels 26 and 27 positioned directly below the lens of the upright microscope, allowing observation of this area. The inlet of flow channel 24 of the microfluidic chip is then connected to the sample injection chamber. When the cell suspension is treated with immunomagnetic beads, the beads bind to the target cells, forming larger cell particles that enter through the inlet of flow channel 24. These particles flow sequentially along flow channel 24 and channel 21. In the sorting channel 21, as fluid flows from the expansion zone to the contraction zone, a secondary flow perpendicular to the main flow is formed, creating two relatively rotating vortices at the inlet of the contraction zone. Due to the velocity gradient, particles of different sizes experience different inertial lifting forces. Larger particles, due to their larger size, experience a stronger inertial lifting force and thus migrate towards side B more quickly. Within the continuous contraction and expansion zones, smaller particles are pushed towards side A, while larger particles move back to side B due to inertial forces, thus achieving spatial separation of particles of different sizes. The target cell particles bound to the magnetic beads then enter flow channel 26 and undergo further sorting through secondary sorting channel 21. Cells located in collection channel 71 are collected for culture.

[0029] To facilitate single-cell collection, the microfluidic channel 2 may optionally include a rinsing channel 25, which is connected to the collection channel 24. Preferably, the rinsing channel 25 is connected to the intersection of the sample inlet channel 24 and the sorting channel 21, thereby forming a three-way structure with the outlet of the rinsing channel 25, the inlet of the sorting channel 21, and the outlet of the sample inlet channel 24 connected together.

[0030] Additionally, it should be noted that the overall size of the microfluidic chip is set according to experimental needs, and this invention does not impose any special limitations. Generally, the microfluidic chip can be set with a width of 20-30mm, a length of 80-150mm, and a thickness of 1-1.5mm. For example, the microfluidic chip has a width of 25mm, a length of 130mm, and a thickness of 1.5mm.

[0031] Although the present invention has been disclosed above, its scope of protection is not limited thereto. Those skilled in the art can make various changes and modifications without departing from the spirit and scope of the present invention, and all such changes and modifications will fall within the scope of protection of the present invention.

Claims

1. A microfluidic chip for skin cell sorting, characterized in that, The base (1) comprising glass material and micro flow channel on the base, the height of the micro flow channel is 30-60um; the micro flow channel comprises 1 primary sorting channel (21) and 2 secondary sorting channels (22, 23); the primary sorting channel (21) is composed of 3 sorting matrices (3) linearly arranged, and the secondary sorting channels (22, 23) are composed of 2 sorting matrices (3) linearly arranged.

2. The microfluidic chip of claim 1, wherein, The sorting matrix (3) is composed of an expansion part (31) connecting a contraction part (32), wherein the width of the contraction part (32) is 30-60um, and the length is 120-200um, the width of the expansion part (31) is 250-500um, and the length is 600-800um.

3. The microfluidic chip of claim 2, wherein, The micro flow channel further comprises a sample inlet channel connected with the primary sorting channel through a sample inlet hole (4), and the width of the sample inlet channel (24) is 300-500um.

4. The microfluidic chip of claim 2, wherein, The micro flow channel further comprises a washing channel (25) connected with the primary sorting channel through a washing hole (5), and the width of the washing channel (25) is 300-500um.

5. The microfluidic chip of claim 3, wherein, The angle between the sample inlet channel (24) and the washing channel (25) is 70-90°.

6. The microfluidic chip of claim 5, wherein, The micro flow channel comprises 2 flow channels (26, 27) connected with the primary sorting channel (21) and the secondary sorting channels (22, 23), wherein the angle between the 2 flow channels (26, 27) is 90-120°.

7. The microfluidic chip of claim 6, wherein, The first secondary sorting channel (22) is connected with 2 first collection holes through 2 collection channels, wherein the included angle between the 2 collection channels is 60-80°.

8. The microfluidic chip of claim 6, wherein, The second secondary sorting channel (23) is connected with 2 second collection holes through 2 collection channels, wherein the included angle between the 2 collection channels is 60-80°.

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