[Histonet] Isopropanol in processing
Matt Ibbs
histol <@t> rat-hole.org
Wed Apr 14 03:15:27 CDT 2004
Dear all,
I have been reading the histonet archives looking for information on
processing through isopropanol instead of ethanol. I've found plenty on
the subject of using isopropanol as a means of removing xylene from the
schedule but hardly anything on using isopropanol as a routine dehydrating
agent.
The head of our department wishes to change our routine processing schedule
as the price of ethanol is always rising and we have problems with taxes
and new customs demands. The answer would appear to be use isopropanol
routinely and we've been trying to get a good working method but find that
either the small biopsies are too brittle or the larger specimens (in
particular slices through material from radical prostatectomy) are
insufficiently processed and fall off the slides.
Presently we're thinking that isopropanol is rather more 'harsh' than
ethanol resulting in the brittleness seen in small biopsies and so we have
tried reducing times in alcohol in an attempt to counteract this. Of
course, that results in the problem of poorly processed larger
specimens. What is the answer? Should we use more short steps with small
increases in concentration (50%, 70%, 80%, 90%, 100%) before going to
xylene or should we use longer incubation times in fewer, stronger
dilutions (70%, 90, 100%, 100%)?
Any suggestions gratefully received. I look forward to hearing your opinions.
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