...how to make cell attached

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Wanderermann
Wanderermann's picture
...how to make cell attached

Hello.

I would like to know how to make recordings in the cell attached configuration. The solution for the pipette is the same that the one for wholle cell or should used the same solution that in the perfusion system? And what to do after made the seal? Should keep the amplifier in the same way (without go to clamp or current) and just increase the gain to see better the currents?

viele Thanks!

Fraser Moss
Fraser Moss's picture
Given you previous posts as

Given you previous posts as well as this one I STRONGLY reccomend that read the following book

Patch Clamping
An Introductory Guide to Patch Clamp Electrophysiology
By Areles Molleman
Wiley Publishers ISBN 0-471-48685-X

Also read the Axon Guide

http://www.moleculardevices.com/pages/instruments/axon_guide.html

For cell attached patch you fill the pipette with the same solution that is bathing the cell (extracellular solution).

!!Remember !! in cell attached mode

1) Positive current = inward membrane current
2) membrane potential = Vrest - Vp

You are going to have to coat your pipettes with Sylgard #184 to keep the noise levels low.

Optimally you will need to use a headstage compatible with your amplifier that has a feedback resistance around 50GOhm. There is a probably a switch on your amplifier that allows you to change the feedback resistance (or capacitance) of your headstage circuit from "whole-cell" mode to "patch" mode.