RE: [Histonet] H and E staining[Scanned]

From:Kemlo Rogerson

Eosin likes Calcium for a reason I can't remember. An old Senior Chief who
taught me always 'mordanted' in calcium formate before putting into aqueous
eosin that contained calcium ions too. I concur over the washing after
lithium as it removes crystals and it's always nice to be clean and manic
rather than dirty and depressed.

Kemlo Rogerson
Cellular Pathology Manager
East Lancashire Hospitals NHS Trust
DD. 01254-294162
Mobile 0774-9754194

-----Original Message-----
From: Weems, Joyce [] 
Sent: 07 December 2004 09:58
To: Ian Ward;
Subject: RE: [Histonet] H and E staining[Scanned]

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.


From: on behalf of Ian Ward
Sent: Tue 12/7/2004 4:24 AM
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

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?



Ian Ward
Senior Technician
E Floor
The Medical School
Tel: 0115 9249924 Ext 36179

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