gluteraldehyde fixation

From:"Gehan, Loralee"


             I need some help with glut fixation.  My experience with
gluteraldehyde is that it is a very very slow fix and it needs to be
refrigerated.  Also I haven't fixed anything in it for more than 24 hours
and the specimens were very small.  Researchers in my lab are insisting that
I fix their very large specimens (adult rabbit spine) in .25% glut in pbs
for about five days.  Then EDTA decal (10%) for two weeks.  I then paraffin
embed these samples or cut frozen sections.   They insist that this will
preserve lac Z enzyme.    
              What are your opinions on this?  All of my experience tells me
that the inner part of the spine will not fix before necrosis sets in.   Has
anyone every done this?  If you have what was your protocol for fixation?
Thanks all in advance. 

Loralee Gehan
University of Rochester


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