3/10/97
cells. The usual glycerol-based mounting media cannot be used because the
dyes "bleed." Molecular Probes recommended something called Cytoseal. Has
anyone tried it? Any other suggestions?
Judy Drazba, Ph.D. (drazbaj@athene.hh.ri.ccf.org)
Confocal Microscopy Facility, NC-3
The Cleveland Clinic Foundation
9500 Euclid Avenue
Cleveland, OH 44195-5001
Office (216)445-3760
FAX (216)444-7927
drazbaj@ATHENE.HH.RI.CCF.ORG
though it is glycerol based, it solidifies in 1 hr or less after mounting. You
could add DABCO or whatever to the glycerin jelly but I don't recall much of a
bleaching problem with DiO.
--
Gordon L. Warren, Ph.D.
Research Scientist
Muscle Biology Laboratory
158 Read Building
Texas A&M University
College Station, TX 77843-4243
Fax (409) 862-4808
Phone (409) 862-4809 office
e-mail root@rangers.tamu.edu
DiO. I have used a mountant that consists of 65% glycerine in PBS pH 7.2 for
the last 6 years. It does NOT cause DiI or DiA to bleed, even over months or
perhaps years depending on the specimens. I section brain tissue using a
vibratome and can often go back to tissue that was sectioned and mounted 1-2
years previously and still get good, photographable results. I cannot speak for
other methods of handling tissue (e.g. cryostat or frozen section which would
fracture the membrane). I have also heard from colleagues that using 100%
glycerine causes the tissue to bleed within weeks.
I have no experience with Cytoseaal mountant.
Hope this helps. Good luck!
ANDREA ELBERGER
Professor of Anatomy and Neurobiology
The University of Tennessee, Memphis
aelberger@utmem1.utmem.edu