[Histonet] H and E staining

[Weems, Joyce]

I would wash after the lithium carbonate.
Is your eosin alcoholic? I would make it in 70% alcohol. Then dip slides in 80% alcohol between the lithium and  eosin. Then go directly into 95% alcohol, absolute, xylene as usual.
Joyce

________________________________

From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Ian Ward
Sent: Tue 12/7/2004 4:24 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] H and E staining



Hi all

I could use some advice, I have been trying to H and E stain some mouse sections of the lungs, brain, spleen, liver, kidney, pancreas and heart. i have used the following method:-
Xlylene     5minutes
Xlylene     5minutes
100% IMS 5minutes
100% IMS 5minutes
90% IMS  5minutes
70% IMS  5minutes
50% IMS  5minutes
running tap water   5minutes
Filtered Haematoxylin   5minutes
Quick wash in running tap water
Dip 20 times in Acid Alcohol
Quick wash in running tap water
Few seconds in Lithium Carbonate
1% Eosin   5minutes
Quick rince in running tap water
Take quickly though alcohols to xylene again
Mount

My problem is that for some reason very little of the Eosin can be seen on the sections when I come to view them. Any ideas?

Cheers

Ian


Ian Ward
Senior Technician
E Floor
The Medical School
QMC
Nottingham
NG7 2UH
Tel: 0115 9249924 Ext 36179


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