From owner-celegans@hgmp.mrc.ac.uk Thu Nov 30 19:49:14 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id E574717A98; Thu, 30 Nov 2000 19:49:12 +0000 (GMT) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 60001) id 99E1517BA4; Thu, 30 Nov 2000 19:49:11 +0000 (GMT) To: celegans@net.bio.net Newsgroups: bionet.celegans Date: Thu, 30 Nov 2000 18:28:56 +0000 From: cbaer@darkwing.uoregon.edu (Charles Baer) Subject: counting worms redux Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Message-Id: <20001130194911.99E1517BA4@mercury.hgmp.mrc.ac.uk> Sender: owner-celegans@hgmp.mrc.ac.uk Precedence: bulk Sirs and Madams, In January I posted a query about digitizing the counting of worms on 60mm agar plates, and I received some helpful replies regarding potential image analysis software (thanks to all who replied). However, it has become apparent that the difficulty is not with image analysis software per se, but rather with the quality of the image we can get with our camera/microscope setup. We are currently using a Diagnostic Instruments RT Color Spot Model 2.2.0 digital camera with the Spot Image grabbing software, and we cannot get sufficiently good images at low magnification (20X) to reliably count all the worms on the plate. The problem to be solved is this: we need to ACCURATELY count dead (or immobilized) worms of all stages (>= L1) on a 60 mm agar plate, and we need very high throughput. A trained observer can currently count by eye all the worms on a plate in approximately 3 minutes. The advantages of automating this process are obvious. If anyone has devised (or can think of) a solution to this particular problem, we will be most grateful if you share it with us. One possibility that has been suggested is a stain that will stain worms but not E. coli. I haven't come across such a stain, but again, suggestions will be most appreciated. Thanks, -Charlie Baer ******************************************************* Charles F. Baer Ecology and Evolution Program 1210 University of Oregon Eugene, OR 97403-1210 USA Phone: 541-346-4697 FAX: 541-346-2364 Email: cbaer@darkwing.uoregon.edu --- .