Re: [Histonet] Novice needs help with vacuum infiltration

From:"Sarah Jones"

Hi Joe,
   Oh, the heady days of processing by hand and Uri Gagarin.  (a little
Hunt for Red October lingo there).  How many baths does your vacuum
infiltrator have?  I assume this is not an autoprocessor, but just for
the paraffins only.  You can put the samples in directly from xylene. 
Can you change the paraffin out with new baths if it only holds one
bath?  How long are you leaving the tissues in paraffin now?  How big
are the samples?  

Sarah Jones HT(ASCP)
Dept. of Vet. Anatomy & Public Health
Histology Lab
Texas A&M University
College Station, TX 77843-4458
phone: 979-845-3177
fax:  979-458-3499

>>> "Joe Amateur"  12/9/2003 3:11:30 PM >>>
Hi all,
I'm a very green, extremely inexperienced novice that has been kind of

thrust into learning histology by chance. It's fascinating stuff and
eager to learn, but the only training I've been given is two textbooks
the Internet. Thus I need help.

We're hand-processing soft (vascular) tissue for H&E in paraffin. We've
a vacuum infiltrator, but no idea how to incorporate it in our
(currently using 3 separate baths of molten paraffin at atmospheric
at 1 hour each). We know that the infiltration time under vacuum is
but can anyone advise what would work best? Could we put the samples in
infiltrator directly out of xylene, or should we incubate the samples
paraffin first to clear out the xylene?

Any help would be deeply, deeply appreciated.

--Thanks in advance,
Jack England

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