[Histonet] Novice needs help with vacuum infiltration

Majid Ghoddusi mghoddusi <@t> cmri.usyd.edu.au
Tue Dec 9 16:50:48 CST 2003

Hi All,

While on this subject, I would appreciate it if you could share your entire
protocol for hand processing with me. Someone in our lab has just started
doing it for his embryo/fetus/muscle samples and he is not having much fun

Majid Ghoddusi

Majid Ghoddusi DVM, PhD
Senior Microscopist
Muscle Development Unit
Children's Medical Research Institute
Locked Bag 23, Wentworthville NSW 2145

-----Original Message-----
From: Sarah Jones [mailto:SJones <@t> cvm.tamu.edu] 
Sent: Wednesday, 10 December 2003 9:35 AM
To: joeamateur <@t> hotmail.com; histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Novice needs help with vacuum infiltration

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

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" <joeamateur <@t> hotmail.com> 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 I'm 
eager to learn, but the only training I've been given is two textbooks and 
the Internet. Thus I need help.

We're hand-processing soft (vascular) tissue for H&E in paraffin. We've got 
a vacuum infiltrator, but no idea how to incorporate it in our protocols 
(currently using 3 separate baths of molten paraffin at atmospheric pressure

at 1 hour each). We know that the infiltration time under vacuum is shorter,

but can anyone advise what would work best? Could we put the samples in the 
infiltrator directly out of xylene, or should we incubate the samples in 
paraffin first to clear out the xylene?

Any help would be deeply, deeply appreciated.

--Thanks in advance,
Jack England

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