[Histonet] vibratome/frozen immuno, soap residue, autofluorescence,
and bad list posting habits
Mike King
making <@t> nersp.nerdc.ufl.edu
Wed Apr 14 12:56:19 CDT 2004
re 9. RE: Vibratome Considerations (Charles Scouten)
...as for immunolabeling sensitivity differences between vibratome and
frozen sections from cryoprotected tissue, is there any science to
support this claim (references)? i've never seen a difference and doubt
there is really any reason to expect one. especially if you need to
permeabilize anyway...
re Danielle Zalinski Vibratome Considerations post:
...50 micron sections of brain (vibratome or other) will label through
the entire thickness depending on the fixation, permeabilization, and
location of the antigen. if you're going to use the tissue for anything
that demands full-thickness labeling, you should use a confocal
microscope to check that the labeling is even through the entire
thickness of the sections. papers in the histo literature will detail
the procedures you need for your material, or post again for specifics.
re 3. autoflourecence in RBC (James Watson)
...try 0.1% sodium borohydride for quenching autofluorescence; this used
to be on the Molecular Probes website. please post your results if this
does/doesn't work.
re 13. Clean Glassware Test (Ostrander, Anita B)
...for testing for soap residue, any bartender worth his/her salt will
tell you that nothing takes the head off a fine beer like a trace of
soap on a poorly rinsed glass. perhaps your lab should stock a few pints
of Guinness?
...and please, please, please, kind histonetters, don't include the
whole digest in replies to histonet. hardly a day goes by when the
digest doesn't contain at least one complete copy of all the previous
day's posts because somebody didn't think before replying. we've all
hit the send button too soon, but this is a real nuisance.
happy histo,
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