Dynamic detection method of potassium ion exchange inside and outside HEK (human embryonic kidney) 293 cell and erythrocyte

Abstract

The invention relates to a simple and easy dynamic detection method of potassium ion exchange inside and outside a human embryonic kidney epithelial cell (HEK 293) and erythrocyte. The method comprises the following steps of: preparing a nano capillary taking a boron silicon capillary as a material, and carrying out inner wall silanization, thus the nano capillary is changed to be hydrophobic; and filling a 1,2-dichloroethane (DCE) solution containing a K<+> vector in the nano capillary with a hydrophobic inner wall tube, wherein the solution belongs to an organic phase, the HEK293 cell and erythrocyte solution belongs to a water phase, Ag / AgCl belongs to the reference electrode in the water phase, Ag / AgTPBCI is a reference electrode in the organic phase. According to the invention, by utilizing a nano tube ion selective micro electrode, the dynamic detection of K<+> exchange inside and outside the HEK 293 cell and erythrocyte is carried out by cyclic voltammetry, thus the perfect combination of the ion selective electrode and an electrochemical detection technology is realized. The method combines the characteristics of a patch clamp technique and the ion selective electrode, and has the advantages of no destruction, low cost, rapid detection and high sensitivity.

Description

technical field [0001] The invention is a simple and highly sensitive new method for dynamic detection of potassium ion exchange inside and outside of HEK 293 cells and erythrocytes, which belongs to the method for electrochemical research on animal cells by using a two-electrode system. Background technique [0002] All living cells are surrounded by a lipid bilayer, or cell membrane, approximately 40 Angstroms thick. The existence of the cell membrane enables various substances in the cell to exist stably and carry out chemical reactions of the living body, and at the same time, it can selectively transport the important substances such as Na + 、K + , Ca 2+ , Cl - wait. The signal transmission in the living system is closely related to the transmembrane transport of ions between cells, and the transport of ions is usually completed by various ion channels on the cell membrane. [0003] The patch clamp technique uses negative feedback electronic circuits to form a h...

AI Technical Summary

Problems solved by technology

Although the patch clamp detection method is widely used in the detection of cell electrophysiology, there are still some shortcomings, such as: relatively poor ion selectivity; certain damage to cells when sealing and breaking the membrane; one time only detect a cell etc.

[0004] Non-destructive micro-measurement technology is used to measure the flow velocity and direction of movement of ions and molecules on the surface of the living body or non-destructively, and does not discuss the ion exchange inside and outside the cell , while the testing equipment of this technology is expensive, which is not conducive to wide application

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