Re: Disappearing Cytoplasm!!

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From:"Tim Morken" <>
Date:Tue, 21 Sep 1999 07:50:00 EDT
Content-Type:text/plain; format=flowed

It sounds like you have some extraction of the cytoplasm. Why not fix 
directly after cutting? There is not really any need to dry so long before 
fixing. You can use ice-cold formalin which can be kept right in the 
cryostat cabinet. Cut and drop the slides in.

Tim Morken, B.A., EMT(MSA), HTL(ASCP)
Infectious Disease Pathology
Centers for Disease Control
1600 Clifton Rd.
Atlanta, GA 30333


Phone: (404) 639-3964
FAX:  (404)639-3043

----Original Message Follows----
From: Bruce Abaloz <>
Subject: Disappearing Cytoplasm!!
Date: Mon, 20 Sep 1999 17:22:03 +1000

Hi all....I'm a student here @ the Uni.of Melbourne & am freezing fresh 
liver & spleen (mouse), floating the OCT covered specimen in a boat in 
liquid nitrogen
until OCT is a solid block then I drop it all into the liquid nitrogen. I 
store it @ -70C until ready to cut in the cryostat.I am using Histogrip 
slides & cut @ 6 microns @ -10C.I let it dry @ room temp.for 1/2 hr.- 2 
hrs.then fix in 10% NBF for 10 min. rinsing X3 in PBS. after which I 
transfer to DH2O to transport to the Histology Lab. for H&E staining. I have 
no problem with the stain but the sections after staining, look as if the 
cytoplasm has been "dehydrated away".Can
anyone advise me on what "to do" or what I'm doing wrong. I've also tried 
Acetone fixation (10 min.@ RT) with the same result??????????? I look 
forward to your replies & THANKYOU in advance.........Simone Cuff.

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