Immunofluorescence

Johnny Fares fares at cuccfa.ccc.columbia.edu
Mon Nov 23 15:13:59 EST 1998


I am using the modified Finney-Ruvkun protocol to prepare worms for
immunofluorescence.  I have no problems with older worms, but I am losing
most of L1-L2 worms during the Borate washes.  Before Borate (with TTE)
the worms spin down to the bottom of the eppendorf tube after one
minute spin at 4000rpm. With the borate, I can see most of my worms stuck
to the side of the tube even after a more vigourous spin.  I tried using
slick ep tubes and that didn't help.  Any suggestions to help me retain my
worms?  I suspect that it tis the detergent in the TTE that is keeping the
worms from sticking to the sides of the tube.  Maybe adding Triton X100 to
the Borate buffer, or is that bad chemistry?

Hanna (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 0383





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