[Histonet] Paraffin Embedding of Mice Tissue - My protocols

[Dallas Nippert]

I worked with mouse tissue for 6 years, and it will almost always be dry.
The only thing I did was soak my blocks in my waterbath for about 45 seconds
after facing, and then put them on my ice.  The protocol for processing was:

70% Etoh 20 min
80% Etoh 20 min
95% Etoh  x2 for 20 min
Absolute  x2 for 20 min
Xylene x2 for 20 min
Paraffin x2 for 30 min
I also used an infiltrating wax in the processor and an embedding wax in the
embedder. The tissues would soak in the wax for about 15 min before I
started and while I was embedding them.
If they are still dry, try cutting down an alcohol time.

Dallas



----- Original Message ----- 
From: "Kelly Salceies" <KSalceies <@t> salud.unm.edu>
To: <Histonet <@t> lists.utsouthwestern.edu>
Sent: Monday, September 29, 2003 11:29 AM
Subject: [Histonet] Paraffin Embedding of Mice Tissue - My protocols


> 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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