70% EtOH, 2 changes, 25 min in each; 95% EtOH, 3 changes, 25 min each; 100% EtOH, 3 changes, 25 min each; xylene, 2 changes, 25 min each; paraffin, 2 changes under vacuum, 61 degrees C using Fisher's Tissue Path Paraplast X-tra paraffin, 45 minutes each.

I have one colleague who runs his mouse colon samples at one hour per station, and they turn out fine, according to him.  (No, I haven't tried this one yet.)

What do you think?
Thanks for the input-
Kathleen
Principal Lab Technician
Neurotoxicology Laboratories
Molecular Pathology Facility Core
Dept of Pharmacology & Toxicology
Ernest Mario School of Pharmacy
Rutgers University
41 B Gordon Rd
Piscataway, NJ 08854
(732) 445-6914
FAX (732) 445-6905

Kemlo Rogerson wrote:

Tissue being too hard is usually due to 'overprocessing' (too long in
the alcohol or clearing agents), paraffin wax too hot or mercury based
fixatives.

Any of these fit?

Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS
Tel: 01270 877625
Mob: 07830 196072
Mobile E-Mail kemlorogerson@3mail.com
FAX & Answer Phone 0871 242 8094
E-mail Accounts:
             kemlo@tiscali.co.uk or
             kemlo1@btinternet.com
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-----Original Message-----
From: Kathleen Roberts (by way of Histonet)
[mailto:kgrobert@rci.rutgers.edu]
Sent: 13 August 2003 12:32
To: HistoNet@pathology.swmed.edu
Subject: Tissue processing problems

To all-

We are a small university pathology lab that does mostly rodent and some

fish tissues, and lately they have been coming out of the processor hard

as a rock and very difficult to cut, and we have been trying to figure
out what, if anything, we are doing wrong.  We have tried changing out
the reagents more often, but it doesn't seem to have made a difference.
We doubt that it's the temperature of the paraffin, as our settings are
in agreement with the manufacturer's recommendations, and the
temperature displayed agrees with our thermometer. Could you please send

me your processing protocols for comparison?

If you like, I will send you our protocol as well.  Any and all ideas
and suggestions will be very welcome.

Thanks in advance for your help-
Kathleen Roberts
Principal Lab Technician
Neurotoxicology Laboratories
Molecular Pathology Facility Core
Dept of Pharmacology & Toxicology
Ernest Mario School of Pharmacy
Rutgers University
41 B Gordon Rd
Piscataway, NJ 08854
(732) 445-6914
FAX (732) 445-6905