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

A dynamic detection, red blood cell technology, applied in the direction of material electrochemical variables, etc., can solve the problems of cell damage, poor ion selectivity, expensive testing equipment, etc., to achieve the effect of small sample volume, high sensitivity and simple operation

Inactive Publication Date: 2012-08-15
NANJING NORMAL UNIVERSITY
View PDF2 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Dynamic detection method of potassium ion exchange inside and outside HEK (human embryonic kidney) 293 cell and erythrocyte
  • Dynamic detection method of potassium ion exchange inside and outside HEK (human embryonic kidney) 293 cell and erythrocyte
  • Dynamic detection method of potassium ion exchange inside and outside HEK (human embryonic kidney) 293 cell and erythrocyte

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] (1) Nano capillary drawing

[0034] The borosilicate capillary (1.0 mm outer diameter, 0.58 mm inner diameter, L = 10 cm) was used as the material to prepare the nanocapillary, and the parameters (Heat, Filament, Velocity, Delay, Pull) of the pulling instrument (P-2000 Puller) were set, and the drawn capillary , to obtain a capillary with a tip diameter ratio of 2.0. (The length of the capillary is 10 cm, and it is broken from the middle to obtain two nanocapillaries with a tip at one end)

[0035] (2) Hydrophobic treatment of the inner wall of the nanocapillary

[0036] Immerse the nanocapillary tip into the trimethylchlorosilane solution for a while (the capillary effect makes the trimethylchlorosilane enter the capillary), and then use a syringe to slowly push the trimethylchlorosilane into the capillary from the back end of the tube to the end of the tube , so that it reaches the tip of the tube, no air bubbles can remain in the capillary and keep it in the tube f...

Embodiment 2

[0051] The borosilicate capillary (1.0 mm outer diameter, 0.58 mm inner diameter, L = 10 cm) was used as the material to prepare nanocapillary. Short capillary (patch-type). The inner surface of the electrode was hydrophobized, and after the electrode was naturally dried, a small amount of DCE solution containing potassium ionophore was injected through a syringe. Then, the organic phase reference electrode Ag / AgTPBCl was inserted into the capillary tube containing the above-mentioned ionophore, and the tip of the assembled electrode and the reference electrode Ag / AgCl in the aqueous phase were respectively inserted into the red blood cell solution (2 mL 2 % red blood cell suspension) below the liquid level, constitute a two-electrode system, connect the device, and obtain the K inside and outside the red blood cell when it is not processed by nanomaterials through cyclic voltammetry test + The exchanged I-V curve, that is, the blank control. Experiments were performed at ro...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

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...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/48
Inventor 赵文波王燕杜春蕾蔡称心黄晓华
Owner NANJING NORMAL UNIVERSITY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap