1. Delayed gramicidin delivery through an intra-pipette capillary facilitates perforated patch recordings.
- Author
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Kazashi K, Miura K, Ueda S, Utsunomiya K, Kiriyama M, Yagisawa M, Ikeda M, Kano M, and Tabata T
- Subjects
- Animals, Neurons physiology, Neurons drug effects, Gramicidin administration & dosage, Gramicidin pharmacology, Patch-Clamp Techniques methods
- Abstract
The gramicidin-perforated patch-clamp technique is indispensable for recording neuronal activities without changing the intracellular Cl
- concentration. Conventionally, gramicidin contained in the pipette fluid is delivered to the cell membrane by passive diffusion. Gramicidin deposited on the pipette orifice sometimes hampers giga-seal formation, and perforation progresses only slowly. These problems may be circumvented by delivering a high concentration of gramicidin from an intra-pipette capillary after a giga-seal is formed. We herein describe the detailed protocol of this improved method. This protocol would greatly facilitate the investigation of Cl- gradient-dependent neuronal activities., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)- Published
- 2024
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