Microfluidic cell processing chip and application method thereof

Abstract

The invention relates to a microfluidic cell processing chip, which comprises a bottom layer and a top layer. The top layer is provided with a serpentine pipe and a cell operation chamber. The solution inlet of the serpentine pipe is provided with two inflow channels. The inlet pipeline is respectively provided with a protective agent inlet and a buffer solution inlet, and the solution outflow channel of the serpentine pipeline is connected to the cell operation chamber, and the cell operation chamber is provided with a solution outlet, a waste liquid outflow channel and a cell inlet and outlet channel, and the cell operation chamber There are a row of cylindrical obstacles on the left and right sides of the inner chamber to confine the cells in the operation chamber and prevent them from being washed away by the protective agent. In actual application, the operator uses a modified cell aspirator to directly blow or suck the cells into or out of the cell operation chamber of the chip. The microfluidic cell processing chip and application method of the present invention can continuously change the concentration of the protective agent when loading or removing the cryoprotectant from larger animal cells, and has the advantages of direct and simple operation and less damage to cells.

Description

technical field [0001] The invention relates to a microfluidic cell processing chip, in particular to a microfluidic chip which can cause the concentration of the cryoprotectant to continuously change during the loading and removal process of the cryoprotectant, and has the advantages of less damage to cells, simple and obvious operation, and the same. application method. Background technique [0002] Cryopreservation of cells is one of the main methods of cell preservation. Using cryopreservation technology to store cells in liquid nitrogen at -196°C can temporarily release the cells from the growth state and preserve their cellular characteristics, so that the cells can be revived for experiments when needed. In addition, proper preservation of a certain amount of cells can prevent cells from being lost due to contamination of the cells being cultured or other unexpected events, and plays a role in cell preservation. When cells are cryopreserved, a protective agent is ad...

AI Technical Summary

Problems solved by technology

The operation steps of this method are cumbersome, and it is required to accurately find the cells under the microscope in a short period of time and complete the transfer of the cells between the solutions, which is very easy to cause the loss of the cells and requires a high level of proficiency for the operator; And the number of step-by-step removals cannot be increased too much, so that the oocytes will still be subjected to several sudden volume changes due to sudden changes in the concentration of the protective agent, and the exposure time to high concentrations of cryoprotectant is long

[0004] Aiming at the above-mentioned problems existing in the prior art, Chinese Patent Publication No. CN103451090A discloses a microfluidic cell processing chip and its application method. It provides a microfluidic cell processing chip that continuously changes the concentration of cryoprotectant. During the removal process, the cryoprotectant flows into the cell operation chamber after mixing through the serpentine channel, and the cells are loaded or sucked out of the chip through the torque control microvalve above the cell inlet and outlet channels, and the cells are restricted by the half partitions on the left and right sides of the operation chamber. In the cell cavity, although the chip and cell processing method can complete the continuous loading and removal of cryoprotectant for large-volume animal cells, there are still many problems in the production and application: (1) the serpentine channel in the chip, the cell The heights of the three important structures of the operating chamber and the half-partition are not uniform. Therefore, in the photolithography stage of processing, photolithography masks need to be layered and made multiple times, and then different layers are bonded together in the sealing stage, which is not very good. The processing accuracy of the guarantee channel and the cavity increases the production cost; (2) the cell controls the opening and closing of the microvalve through the torque to control the cell entering and exiting the cell operation cavity, and the switch state of the valve is controlled by the screw fixed above the channel of the PDMS chip , the deformation part between the screw end and the fluid channel is a thin layer of polyurethane material. This design requires the coupling of two materials, PDMS and polyurethane, on the chip, which increases the complexity of chip processing. Although this microvalve has a simple structure, it does not require It is a complex control system, but it is difficult to automate in the later stage, and it is only suitable for simple microfluidic systems and laboratory operations; (3) half partitions are used on the left and right sides of the operation chamber to limit cells, and the height of the channel at the half partition can be adjusted according to the cell However, the height of the channel at the semi-partition is significantly smaller than that of the serpentine channel, and the resistance of the liquid flowing through it will increase, and the pressure drop value of the system will increase, which will easily cause damage to the microfluidic chip and shorten the Chip service life

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