A method to improve the efficiency of patch clamp experiment

A technology of experimental efficiency and patch clamping, applied in the direction of biochemical equipment and methods, microorganisms, animal cells, etc., can solve the problems of low experimental efficiency, high operation difficulty, affecting the effect of traditional patch clamping, etc., to increase the adhesion strength , improve transfection efficiency, and increase the average cell survival time

Active Publication Date: 2022-05-13
广州浚远康生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] To sum up, the traditional patch clamp is the most important experimental technique in the field of electrophysiology, but due to the extremely difficult operation and low experimental efficiency, it seriously affects the traditional patch clamp to fully play its role.

Method used

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  • A method to improve the efficiency of patch clamp experiment
  • A method to improve the efficiency of patch clamp experiment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 A method for maintaining the state of cells to improve the efficiency of patch clamp experiments

[0029] S1, 6-well cell culture plate coating:

[0030] Coating of 6-well cell culture plate: Add 100 μL of Matrigel (Matrigel) to 10 mL of serum-free DMEM / F12 medium at 4 °C (diluted 100 times), mix well to obtain a mixture, and use a pipette gun to Pipette the above mixture into a 6-well cell culture plate, 1 mL per well, and place the 6-well cell culture plate in CO 2 In a cell culture incubator, 37°C, 5% CO 2 , and cultured statically for 1 h to obtain a Matrigel-coated 6-well cell culture plate, which is set aside.

[0031] S2. Cell culture:

[0032] Cell recovery: Take out a cryopreservation tube containing 1mL HEK293T cells from the liquid nitrogen tank, clamp the cryopreservation tube with tweezers and put it in a 37°C water bath to dissolve the cells quickly; then add the cells to 8mL containing 10% In DMEM medium containing fetal bovine serum, centri...

Embodiment 2

[0038] Example 2 A method for maintaining the state of cells to improve the efficiency of patch clamp experiments

[0039] S1, 6-well cell culture plate coating:

[0040] Coating of 6-well cell culture plate: Add 100 μL of Matrigel (Matrigel) to 15 mL of serum-free DMEM / F12 medium at 4 °C (diluted 150 times), mix well to obtain a mixture, and use a pipette gun to Pipette the above mixture into a 6-well cell culture plate, 1 mL per well, and place the 6-well cell culture plate in CO 2 In a cell culture incubator, 37°C, 5% CO 2 , and cultured statically for 1 h to obtain a Matrigel-coated 6-well cell culture plate, which is set aside.

[0041] S2. Cell culture:

[0042] Cell recovery: Take out a cryopreservation tube containing 1mL HEK293T cells from the liquid nitrogen tank, clamp the cryopreservation tube with tweezers and put it in a 37°C water bath to dissolve the cells quickly; then add the cells to 8mL containing 10% In DMEM medium containing fetal bovine serum, centri...

Embodiment 3

[0048] Example 3 A method for maintaining the state of cells to improve the efficiency of patch clamp experiments

[0049] S1, 6-well cell culture plate coating:

[0050] Coating of 6-well cell culture plate: Add 100 μL of Matrigel (Matrigel) to 20 mL of serum-free DMEM / F12 medium at 4°C (diluted 200 times), mix well to obtain a mixture, and use a pipette gun to Pipette the above mixture into a 6-well cell culture plate, 1 mL per well, and place the 6-well cell culture plate in CO 2 In a cell culture incubator, 37°C, 5% CO 2 , and cultured statically for 1 h to obtain a Matrigel-coated 6-well cell culture plate, which is set aside.

[0051] S2. Cell culture:

[0052] Cell recovery: Take out a cryopreservation tube containing 1mL HEK293T cells from the liquid nitrogen tank, clamp the cryopreservation tube with tweezers and put it in a 37°C water bath to dissolve the cells quickly; then add the cells to 8mL containing 10% fetal In DMEM medium with bovine serum, centrifuge at...

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Abstract

The invention discloses a method for improving the efficiency of a patch clamp experiment, comprising the following steps: coating a cell culture plate with Matrigel to obtain the Matrigel-coated cell culture plate, subculture the cells after recovery, and subculture the subcultured Cells were seeded into Matrigel-coated cell culture plates for culture, and then DNA or plasmids were transfected into the cells, and the transfected cells were added to glass slides for patch clamp experiments. The method of the invention increases the average survival time of cells by 200%-400% when loading samples, and significantly improves the efficiency of the patch clamp experiment.

Description

Technical field: [0001] The invention belongs to the technical field of electrophysiological patch clamp, and in particular relates to a method for improving the efficiency of patch clamp experiments. Background technique: [0002] The patch clamp technique is a technique that records the ion current passing through the ion channel to reflect the molecular activity of the ion channel on the cell membrane. It is an electrophysiological experimental technique used to study the ion flow of a single isolated living cell, tissue slice or cell membrane. This technique plays a crucial role in the study of excitable cells such as neurons, cardiomyocytes, myofibers and pancreatic cells, and can also be used to study bacterial ion channels in specially prepared giant spheroids. [0003] The traditional patch clamp technique has very high technical requirements for the experimenters. Generally, the experimenters need to undergo strict and long-term training in order to operate accurat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/07
CPCC12N5/06C12N2533/90
Inventor 李志远汤凤徐浚杰田超刘雨杰黄荣奇
Owner 广州浚远康生物科技有限公司
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