PMID: 7539128Jan 1, 1995Paper

Improved micro-perfusion chamber for multiple and rapid solution exchange in adherent single cells

Pflügers Archiv : European journal of physiology
A SavchenkoS Hering

Abstract

A new method for fast perfusion of single adherent cells during whole-cell and single-channel patch-clamp measurements is described. The main advantages over previous methods are: (1) the solution surrounding a single cell in a Petri dish (or cover slip) can be exchanged without contamination of the entire cell population, and (2) only small quantities of test solution (50 microliters) are required. The method consists of inserting a patch pipette into a micro-perfusion chamber (MPC) where whole-cell and single-channel currents can be studied. A small volume (2-3 microliters) surrounding adherent single cells in a Petri dish is separated by enclosing it in a ring of Silicone rubber which is then pressed to the bottom of the dish. A drug-containing test solution (50 microliters) is supplied to a funnel at the chamber inflow. The MPC volume isolated from the main solution in the Petri dish is rapidly changed when suction is applied to the chamber outflow. The speed of solution exchange in the MPC (16 +/- 5 ms, n = 12) was estimated by observing changes in the tip potential of 1-M omega patch pipettes during rapid chamber perfusion with solutions of different ionic composition. The seal of the MPC with the surface of a Petri dish ...Continue Reading

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Citations

Jun 1, 1996·British Journal of Pharmacology·S BerjukowS Hering
Nov 1, 1996·British Journal of Pharmacology·S BerjukovJ Striessnig
Feb 10, 2010·IEEE Transactions on Nanobioscience·Akira YamadaKeiji Naruse

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