From owner-biophys@hgmp.mrc.ac.uk Sat Sep 8 16:09:47 2001 Return-Path: Received: from localhost (localhost [127.0.0.1]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id E6CEE17AD2 for ; Sat, 8 Sep 2001 16:09:45 +0100 (BST) Received: from localhost (localhost [127.0.0.1]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id E068A17B4D; Sat, 8 Sep 2001 16:09:39 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 555F617B23; Sat, 8 Sep 2001 16:09:34 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id QAA14658; Sat, 8 Sep 2001 16:07:55 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Dominic-Luc Webb molmed X-Newsgroups: bionet.biophysics,bionet.neuroscience Subject: Re: Perforated Patch Problem Date: Sat, 8 Sep 2001 17:07:44 +0200 Organization: Stockholm University Lines: 22 Message-ID: References: Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII X-Complaints-To: abuse@su.se To: Matthew Ireland In-Reply-To: To: biophys@hgmp.mrc.ac.uk.neur-sci@hgmp.mrc.ac.uk Sender: owner-biophys@hgmp.mrc.ac.uk Precedence: bulk On 3 Sep 2001, Matthew Ireland wrote: > Dear Colleagues > > Currently I am using perforated patch on neurons within the CNS. > Recently, I have been experinceing a problem when I try to perforate a > neuron, instead of a full perforation as you would expect, it only > results in a partial perforation. Not sure what a partial perforation is. Are you in fact getting a proper seal? I suppose the ampB could go bad, or it could leak out of the pipette and permeabilize the cells as well. Are you back filling a small volume of the ampB into a large volume of ampB-free buffer in the pipette? Cheers, Dominic .