Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Formation method of inside-out patch in patch clamp experiment

An inside-out, experimental method technology, applied in the field of inside-out patch formation of the patch clamp, to achieve the effect of improving experimental efficiency, simple operation, and increasing success rate

Inactive Publication Date: 2013-11-20
LUZHOU MEDICAL COLLEGE
View PDF0 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to overcome the problem that the free cell membrane at the tip of the glass microelectrode will automatically fuse to form vesicles during the inside-out mode patch clamp experiment in the prior art, and the vesicles cannot be ruptured to form a single-layer membrane after being processed by a conventional scheme. To provide an experimental method to quickly and effectively eliminate vesicles and form an inside-out membrane

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
  • Formation method of inside-out patch in patch clamp experiment
  • Formation method of inside-out patch in patch clamp experiment
  • Formation method of inside-out patch in patch clamp experiment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0031] Hereinafter, taking HEK-293 cells recombinantly expressing the large conductance calcium-activated potassium channel (BKCa) of human vascular smooth muscle as an example, the operation method and steps of the present invention will be described in detail. However, it should not be understood that the scope of the above subject matter of the present invention is limited to the following embodiments, and all technologies realized based on the content of the present invention belong to the scope of the present invention.

[0032] 1. Liquid preparation

[0033] Electrode solution: D-aspartic acid40mM, KOH40mM, KCl100mM, HEPES10mM, CaCl 2 1mM.

[0034] Low calcium bath: D-aspartic acid100mM, KOH100mM, KCl40mM, HEPES10mM, EGTA1mM, free Ca 2+ Concentration 10 -5 mM.

[0035] High calcium bath: D-aspartic acid100mM, KOH100mM, KCl40mM, HEPES10mM, EGTA1mM, free Ca 2+ Concentration 10 -4 mM.

[0036] Add 10mM Glucose to all liquids, adjust the osmotic pressure to 284mOSM, a...

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 discloses a formation method of an inside-out patch in a patch clamp experiment. The method comprises following steps: carrying out an inside-out patch clamp experiment according to conventional technology means, when detecting the decreasing or disappearance of the current signal during the current detection process, lifting the electrode out of the bath liquid to expose the electrode in the air, blowing at the tip of the electrode with an air flow with a constant pressure and an even flow velocity, and soaking the electrode in the bath liquid after the blowing is over to eliminate vesicles so as to obtain an inside-out patch. The formation method is simple and convenient, is capable of cracking vesicles formed in the tip of the electrode to form a single-layer patch, is especially suitable for the situations that easy formation of vesicles and difficult rupture of vesicles because of stable vesicle structure are easy to occur in the inside-out formation of the patch clamp technology, can overcome the defect that vesicles are easy to form on the tip of the electrode in a inside-out patch clamp experiment, and thus improves the success rate of the experiment.

Description

technical field [0001] The invention relates to an improved method for a cell patch clamp experiment, in particular to a method for forming an inside-out patch of a patch clamp, and belongs to the field of biological cell experiments. Background technique [0002] The inside-out mode, that is, the inside-out mode, is an important and commonly used method in patch clamp experiments. In this configuration, the inner side of the cell membrane is exposed to the experimental bath, by displacing the bath or directly in the bath. Adding test substances into the liquid can easily change the concentration of ions or substances inside the cell, and can also directly add enzymes or drugs to the inner surface of the membrane. Necessary means of current characteristic influence. [0003] In the process of forming the inside-out configuration of the cell membrane, first contact the cell membrane with a microtube glass electrode, and form a high-resistance seal through negative pressure s...

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): C12N13/00
Inventor 杨艳曾晓荣毛亮程俊文静黄文俊周文
Owner LUZHOU MEDICAL COLLEGE
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products