[Histonet] Exploding skin samples

[Tony Henwood]

Margaret,

Your processing times are too short. 
When the block is faced can you smell xylene from it?
Your processing schedule seems Oh so difficult and complex! Keep it simple.
Refer to a good Histotechnology book.


Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager
The Children's Hospital at Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: 612 9845 3306
Fax: 612 9845 3318


-----Original Message-----
From: Margaret Blount [mailto:mab70 <@t> medschl.cam.ac.uk] 
Sent: Tuesday, 22 June 2004 2:27 AM
To: Histonet (E-mail)
Subject: [Histonet] Exploding skin samples


I have been given some mouse skin fixed in formalin (neutral buffered from
Sigma, equivalent to what we used to call 10% neutral buffered formalin). I
have tried several processes and even tried post fixing in Bouin's fluid as
suggested by a former colleague who did a lot of rat skin sections in a
previous job. They are spreading on the waterbath, especially around the fat
layer at the base of the dermis. It is important to us to get good sections
of these samples, can anyone suggest any other procedures I can try? My
process times were 30 minutes in 50, 60, 70, 80, 90., 100% ethanol, one
ethanol for 1 hour, 50:50 ethanol/xylene 30 mins, ethanol 30 mins, 2
xylenes 30 mins, 2 x paraffin for 30 mins, embed. I only have 2 wax baths on
my processor and have tried returning samples to wax for longer impregnation
times without success. 

I propose to try out a number of different fixatives such as formol alcohol,
methacarn, Bouin's as primary fixation and compare with NBF. Any suggestions
would be most gratefully received.

Thanks a lot

Margaret

Margaret Blount
Chief Technician
Clinical Biochemistry
University of Cambridge
Addenbrooke's Hospital
Hills Road
Cambridge
CB2 2QR 


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