[Histonet] Paraffin Embedding of Mice Tissue - My
protocols
Sarah Jones
SJones <@t> cvm.tamu.edu
Mon Sep 29 16:45:53 CDT 2003
Thanks for the info Kelly, I would use Pro-par or Shandon's xylene
substitute instead of xylene. Your times look ok, although I don't know
what size of your samples. Two absolutes should be enough. Check the
temp of your paraffin, I use the lowest melting point paraffin possible
on the processor and make sure the processor is programmed to be just
2-4 degrees over the melting point. Also, check the temperature with a
thermometer once just to make sure.
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
>>> "Kelly Salceies" <KSalceies <@t> salud.unm.edu> 09/29/03 11:29AM >>>
Hi --
I guess I should have mentioned the protocols I have already tried -
sorry...I'm new to this Histonet thing :)
On the Shandon Hypercenter XP I've tried:
80% ETOH 5 min
95% ETOH 15 min 2X
100% ETOH 15 min 3X
Xylene 10 min 2X
Xylene 5 min 2X
Paraffin 15 min 2X
50% ETOH 5 min
80% ETOH 5 min
95% ETOH 15 min 2X
100% ETOH 15 min 3X
Xylene 10 min 2X
Xylene 5 min
Paraffin 15 min 2X
80% ETOH 5 min
95% ETOH 5 min 2X
100% ETOH 5 min 2X
1:1 ETOH: Cedarwood Oil 5 min
1:1 Cedarwood Oil:Methyl Salicylate 5 min
100% Methyl Salicylate 5 min
1:1 Methyl Salicylate:Xylene 5 min
Xylene 5min
Paraffin 15 min 2X
--The last protocol left the tissue soft - so I might just need to use
longer times...I'm not sure if anybody uses this protocol....
On the Microwave I've tried:
100% ETOH 10 min 2X
1:1 ETOH:Xylene 10 min
Paraffin 10 min
-All steps: Max temp 65 degrees - Watt=750
100% ETOH 5 min
IPA 5 min
Paraffin 5 min
-All steps: Max temp 65 degrees - Watt=750
100% ETOH 5 min
1:1 IPA:Xylene 5 min
Paraffin 5 min
-All steps: Max temp 65 degrees - Watt=750
There are the protocols I've tried....Please let me know what
suggestions you have - thank you!!
Kelly Salceies
University of New Mexico
Health Sciences Center
Dept. of Pathology
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