Re: [Histonet] Fixation for lectin work

From:Gayle Callis


Formalin works.  Have done frozen sections using acetone/alcohol, acetone
fixation and these work also.  If you do formalin or paraformaldehyde
fixation, use trypsin digestion.  The only problem I can see with lectins
and formalin is immunofluorescent staining and the autofluorescence created
by aldehyde fixation. I also have a wonderful review of autofluorescence
(although they wrote it for GFP), it still tells sources of this problem
along with fixation considerations and still useful for other fluorescent
technics.  If you want, I will be glad to attach to you privately.  

There is a wonderful book on Lectin Immunohistochemistry from
Springer-Verlag, very cheap that deals with all lectins and tells what they
stain (more than one thing is not uncommon).   There are buffer
considerations - avoiding PBS with certain lectins may be necessary and you
will have to use the Lectin Buffer TRIS with Ca and Mg added.  

Also, E-Y Laboratories puts out a freebie Lectin book, with list of all
lectins and their inhibiting sugars, as does Vector.  Vector catalog
supplies the concentration on inhibition sugar with lectin to create your
negative control.    

Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology 
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)

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