Formation method of inside-out patch in patch clamp experiment

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...

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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

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