From owner-celegans@net.bio.net Sun Apr 27 23:00:00 1997 Path: biosci!fcs280s.ncifcrf.gov!cpk-news-feed1.bbnplanet.com!cpk-news-hub1.bbnplanet.com!news.bbnplanet.com!news.maxwell.syr.edu!news.new-york.net!news.columbia.edu!cuccfa.ccc.columbia.edu!fares From: Johnny Fares Newsgroups: bionet.celegans Subject: Anesthetize GFP: Methods Date: Mon, 28 Apr 1997 11:51:48 -0400 Organization: Columbia University Lines: 47 Message-ID: NNTP-Posting-Host: cuccfa.ccc.columbia.edu Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII I got a lot of responses for different ways to anesthetize worms for viewing GFP. I will give a brief summary of the differnt methods that were proposed and will put in parantheses the name of the person who suggested the particular treatment (for further information). I had asked about these methods for the purpose of laser ablating cells that had been marked by GFP. Therefore, the worms had to be able to recover from the treatment and I will indicate what is known concerning that. Thank you very much for all your responses. 1) 1mM Levamissole: Worms do recover, but is not known if 100% (M. Stern lab, Rob Steven). 2) 25 mM Azide: in the agar pad and used routinely for GFP-ablations (Theresa Rakow). 3) 1% Nicotine: in S buffer. Recovery? (John Thaden) 4) Ethanol: -750 mM. Reversed in 15 minutes. (Philip Morgan) -8% (1739 mM) in M9 buffer. (David Hall) 5) Chloroform: 2% in air (in closed container): reversed in 30 minutes (Philip Morgan). 6) Ether: 6% in air (in closed container). (Philip Morgan) 7) CO2: -contact Wayne Forrester in Gian Garriga's lab. (Bruce Wightman) -put in sealed box with dry ice for 5 minutes. (Jeff Yuan) -blow on worms from an aspirator filled with dry ice. (Mona Spector) 8) cold shock: I gather this was not done btu was suggested as an option. (Jon Pierce, David Hall). 9) Use paralyzed UNC: This would of course preclude the identification of UNC as a phenotype after ablation. (Jane Hubbard) Johnny Fares 701 West 168 Street HHSC Room 718 Dept. of Biochemistry Columbia University New York, NY, 10032 Tel: 212 305 6931 Fax: 212 305 1721 .