RE: [Histonet] myelin stain for 40um cryostat sections

From:Kemlo Rogerson

Myelin can be stained with Sudan 3 and iron haematoxylin.

Cut frozen (10 U) sections and chromate in 2.5% potassium dichromate for 2
to 4 days, then wash.

5ml fresh 1% aqueous haematoxylin and 5 ml 4% iron alum fresh in a covered
dish. Put your frozen sections in for 45 min at 60 degrees C. Agitate
gently. Wash in water.

Decolorise in 0.5% iron alum for 1 hr with agitation.

Wash in water

Treat with 1% borax/ 2.5% potassium ferricyanide solution for 10 min with

Wash in water

6mls stock Sudan 2 (CI 12140) saturated in 99% isopropanol and dilute with 4
ml water, let stand 7 to 8 min and filter. Stain sections 10 min.

wash in water

Float onto slides, drain and mount in aqueous mountant.

This is from Lillie and I used it decades ago with success; but the old ways
are usually the best.

Kemlo Rogerson
Pathology Manager
Ext  3311
DD   01934 647057
Mob 07749 754194

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-----Original Message-----
From: Maria Mejia [] 
Sent: Wednesday, March 08, 2006 3:19 PM
Subject: [Histonet] myelin stain for 40um cryostat sections

Could someone please recommend a good myelin stain protocol for 40um  
primate brain sections (both for fixed and fresh frozen sections? 10%  
NBF has been
used on the fixed sections. I would greatly appreciate any  
information you could
provide & thank you in advance.

Maria Bartola Mejia
University California San Francisco (UCSF)
Department of Neurosurgery
San Francisco, CA 94103

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