Anesthetize GFP: Methods

Johnny Fares fares at cuccfa.ccc.columbia.edu
Mon Apr 28 10:51:48 EST 1997


     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





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