[Histonet] Paraffin Embedding of Mice Tissue - My protocols

[Sarah Jones]

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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