[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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