Micro-fluidic chip and method for cell sorting and positioning
阅读说明：本技术 一种细胞分选与定位的微流控芯片及方法 (Micro-fluidic chip and method for cell sorting and positioning ) 是由 靳欣 李歧强 韩琳 王兴元 于 2019-09-12 设计创作，主要内容包括：本发明公开了一种细胞分选与定位的微流控芯片及方法,该芯片包括主流道、分选流道和压力控制流道,主流道一端为样品注射口和鞘液注射口,另一端为主流道出口；分选流道和压力控制流道一端密封,另一端分别为分选流道出口和压力控制流道出口；主流道和分选流道之间通过数个连接流道连接,分选流道和压力控制流道之间通过数个微流道连接；分选流道与微流道连接处为碗状结构；分选流道和压力控制流道设置多组,沿着主流道的样品流动方向依次分布于主流道的不同位置,且每组分选流道与主流道之间的连接流道的直径逐级增大,本发明所公开的芯片聚焦精度高,成本低廉,实验步骤简单,对于海洋生态环境研究以及动物组织或血液样品检测预处理非常有意义。(The invention discloses a micro-fluidic chip for cell sorting and positioning and a method thereof, wherein the chip comprises a main flow channel, a sorting flow channel and a pressure control flow channel, wherein one end of the main flow channel is provided with a sample injection port and a sheath fluid injection port, and the other end of the main flow channel is provided with a main flow channel outlet; one end of the separation flow channel and one end of the pressure control flow channel are sealed, and the other end of the separation flow channel and the other end of the pressure control flow channel are respectively a separation flow channel outlet and a pressure control flow channel outlet; the main flow channel is connected with the separation flow channel through a plurality of connecting flow channels, and the separation flow channel is connected with the pressure control flow channel through a plurality of micro flow channels; the joint of the sorting flow channel and the micro flow channel is of a bowl-shaped structure; the chip disclosed by the invention has the advantages of high focusing precision, low cost and simple experimental steps, and is very significant for marine ecological environment research and animal tissue or blood sample detection pretreatment.)
1. A micro-fluidic chip for cell sorting and positioning is characterized by comprising a main flow channel, a sorting flow channel and a pressure control flow channel, wherein one end of the main flow channel is provided with a sample injection port and a sheath fluid injection port, and the other end of the main flow channel is provided with a main flow channel outlet; one end of the separation flow channel and one end of the pressure control flow channel are sealed, and the other end of the separation flow channel and the other end of the pressure control flow channel are respectively a separation flow channel outlet and a pressure control flow channel outlet; the main flow channel is connected with the separation flow channel through a plurality of connecting flow channels, and the separation flow channel is connected with the pressure control flow channel through a plurality of micro flow channels; the joint of the sorting flow channel and the micro flow channel is of a bowl-shaped structure; select separately the runner and the pressure control runner sets up the multiunit, distributes in the different positions of sprue in proper order along the sample flow direction of sprue, and the diameter that every group selects separately the connection runner between runner and the sprue increases step by step.
2. The microfluidic chip for cell sorting and positioning according to claim 1, wherein the main channel is an S-shaped channel comprising a horizontal section and a curved section, and the sorting channel and the pressure control channel are located outside the horizontal section of the S-shape and are parallel to the horizontal section.
3. The microfluidic chip for cell sorting and positioning according to claim 1, wherein the diameter of the micro flow channel is much smaller than the diameter of the cell.
4. The microfluidic chip for cell sorting and positioning according to claim 1, wherein the sorting channel outlet and the pressure control channel outlet are located at two opposite ends.
5. The microfluidic chip for cell sorting and positioning according to any one of claims 1-4, wherein the chip is made of PDMS, PMMA, silicon or glass.
6. A method for cell sorting and positioning using the microfluidic chip for cell sorting and positioning according to claim 1, comprising the steps of:
respectively injecting a sample solution containing cells and a sheath liquid from a sample injection port and a sheath liquid injection port, extracting the liquid from an outlet of a first group of pressure control flow channels, enabling a batch of cells with the smallest size in the sample solution to enter a sorting flow channel through a connecting flow channel and enter a bowl-shaped structure, and then sequentially opening an outlet of a pressure control flow channel of a rear group and extracting the liquid, so that the cells in the sample solution are sorted out in a small-to-large order and are positioned in the bowl-shaped structure; and then carrying out image detection, finally pressurizing through a pressure control flow passage outlet, extracting liquid from a sorting flow passage outlet, and extracting cells positioned in the bowl-shaped structure from different sorting flow passages so as to obtain the cells in a certain size interval, thereby facilitating the detection and analysis of the next step.
Technical Field
The invention relates to a microfluidic technology, in particular to a microfluidic chip and a method for cell sorting and positioning.
Background
In the research of marine ecological environment, microalgae is used as a main energy source of coastal water areas and has important influence on offshore environment. Through the identification of fluorescence and images, the species, the propagation and the biomass of the microalgae can be obtained, so that the water body environment is evaluated and predicted. The separation and observation of the biological tissue and the blood sample also have great significance in scientific research and disease diagnosis and treatment.
The focusing of the microscope and the overlapping of the cells are major challenges for image acquisition and fluorescence identification. By reducing the sample concentration, the problem of cell image overlap can be alleviated to some extent. However, a large number of blank pictures are brought, and the detection efficiency and the post-image processing are adversely affected. The best method for solving the problem at present is to use a flow cytometer, but the flow cytometer is expensive, bulky, inconvenient to carry, complex in detection steps and high in detection cost.
Disclosure of Invention
In order to solve the technical problems, the invention provides a micro-fluidic chip for cell sorting and positioning and a method thereof, so as to achieve the purposes of effectively improving the problems of low focusing precision, overlapping of cell images and the like, low cost and simple experimental steps.
In order to achieve the purpose, the technical scheme of the invention is as follows:
a micro-fluidic chip for cell sorting and positioning comprises a main flow channel, a sorting flow channel and a pressure control flow channel, wherein one end of the main flow channel is provided with a sample injection port and a sheath fluid injection port, and the other end of the main flow channel is provided with a main flow channel outlet; one end of the separation flow channel and one end of the pressure control flow channel are sealed, and the other end of the separation flow channel and the other end of the pressure control flow channel are respectively a separation flow channel outlet and a pressure control flow channel outlet; the main flow channel is connected with the separation flow channel through a plurality of connecting flow channels, and the separation flow channel is connected with the pressure control flow channel through a plurality of micro flow channels; the joint of the sorting flow channel and the micro flow channel is of a bowl-shaped structure; select separately the runner and the pressure control runner sets up the multiunit, distributes in the different positions of sprue in proper order along the sample flow direction of sprue, and the diameter that every group selects separately the connection runner between runner and the sprue increases step by step.
In the above-mentioned scheme, the S type runner that the sprue comprises horizontal segment and crooked section, select separately the runner and the pressure control runner is located the horizontal segment outside of S type, and is parallel with the horizontal segment.
In the above embodiment, the diameter of the micro flow channel is much smaller than the diameter of the cell.
In the above scheme, the sorting flow channel outlet and the pressure control flow channel outlet are located at two ends relatively far away.
In a further technical scheme, the chip is made of PDMS, PMMA, silicon or glass.
A method for sorting and positioning cells adopts the microfluidic chip for sorting and positioning cells, and comprises the following processes:
respectively injecting a sample solution containing cells and a sheath liquid from a sample injection port and a sheath liquid injection port, extracting the liquid from an outlet of a first group of pressure control flow channels, enabling a batch of cells with the smallest size in the sample solution to enter a sorting flow channel through a connecting flow channel and enter a bowl-shaped structure, and then sequentially opening an outlet of a pressure control flow channel of a rear group and extracting the liquid, so that the cells in the sample solution are sorted out in a small-to-large order and are positioned in the bowl-shaped structure; and then carrying out image detection, finally pressurizing through a pressure control flow passage outlet, extracting liquid from a sorting flow passage outlet, and extracting cells positioned in the bowl-shaped structure from different sorting flow passages so as to obtain the cells in a certain size interval, thereby facilitating the detection and analysis of the next step.
According to the technical scheme, the micro-fluidic chip for cell sorting and positioning is connected with the main flow channel and the sorting flow channel through connecting flow channels with different diameters, so that cells with different sizes sequentially enter different sorting flow channels and are positioned in the bowl-shaped structure, the pressure in the bowl-shaped structure is controlled by the pressure control flow channel, the cells can be positioned in the bowl-shaped structure, image detection is facilitated, the focusing precision of a microscope can be effectively improved, the problem of cell superposition is thoroughly solved, the cells to be identified are arranged in order, and the accuracy of pattern identification can be greatly improved.
Meanwhile, the S-shaped main flow channel is adopted, the flow channel area is small, and the photoresist stress can be reduced compared with a large-area region to be sorted, so that the manufacturing yield is improved. Adopt two liquid inlets and S type sprue that sample injection port and sheath liquid injection port constitute, utilize laminar flow and centrifugal force, can make most of cells along the sprue outside motion, and then make most of cells within the biggest size all can be screened the location.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below.
FIG. 1 is a schematic diagram of an internal structure of a microfluidic chip for cell sorting and positioning according to an embodiment of the present invention;
fig. 2 is a partially enlarged schematic view of the embodiment of the present invention.
In the figure, 1, a main flow channel; 2. a sorting flow channel; 3. a pressure control flow path; 4. a sample injection port; 5. a sheath fluid injection port; 6. an outlet of the main runner; 7. an outlet of the sorting flow channel; 8. a pressure control flow passage outlet; 9. connecting the flow channel; 10. a micro flow channel; 11. a bowl-shaped structure; 12. a cell.
Detailed Description
The technical solution in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention.
The invention provides a micro-fluidic chip for cell sorting and positioning and a method thereof, and the chip can effectively improve the cell image acquisition quality and has great significance for marine ecological environment research and detection pretreatment of animal samples such as animal tissues or blood samples and the like, such as a structure shown in figure 1.
The micro-fluidic chip for cell sorting and positioning as shown in fig. 1 comprises a
The
The
The S-shaped flow channel designed by the invention can make the cell solution deflect to the outer side of the S-shaped flow channel under the action of laminar flow, and is convenient to enter the
The micro-fluidic chip designed by the invention has high structural strength, so that the micro-fluidic chip has no excessive requirements on materials, and can be made of various materials, such as PDMS, PMMA, silicon or glass.
A method for sorting and positioning cells adopts the microfluidic chip for sorting and positioning cells, and comprises the following processes:
firstly, respectively injecting a sample solution containing cells and a sheath solution from a sample injection port 4 and a sheath
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
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