Re: [Histonet] processing fatty tissue

From:Maxim Peshkov



Robert:
In my protocol I used graduate alcohols
(isopropanol) for dehydratation, then mixtures
isopropanol:mineral oil 5:1 and 1:2 at 50oC, then
mineral oil instead xylene. Paraffin's
infiltration as usual.
These reagents give the "soft" transitions from
dehydratant to paraffin. Difficult tissues will
cut more easily. Risk of inhalations toxic
chemicals decreased to 5 times.
If I had VIP processor and 100% ethanol, that
I simple used Rene's protocol.

My protocol based on protocol Buesa
(RJ Buesa, JOH/Vol.23, 2/June 2000. Mineral Oil:
The Best Xylene Substiute for Tissue Processing
Yet?).

1 Isopropanol @70%  1h RT
2 Isopropanol @80%  1h RT
3 Isopropanol @90%  1h RT
4 Isopropanol @92%  1h RT
5 Isopropanol 99% (fresh) 1h RT
6 IM 5:1 1.5h 50
7 IM 4:2 1.5h 50
8 Mineral oil 50 overnight
9 Paraffin 1h 60
10 Paraffin 1h 60
11 Paraffin 1h 60
12 Paraffin 1h 60

Now we routinely process all surgical specimens
include bone, eyeball, uterus etc with it.
I hope this protocol would like to you.

Sincerely,
Maxim Peshkov
Russia
Taganrog.


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