[Histonet] myelin stain for 40um cryostat sections
kemlo.rogerson <@t> waht.swest.nhs.uk
Wed Mar 8 09:35:53 CST 2006
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.
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From: Maria Mejia [mailto:mbmphoto <@t> gmail.com]
Sent: Wednesday, March 08, 2006 3:19 PM
To: histonet <@t> lists.utsouthwestern.edu
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
Email: mbmphoto <@t> gmail.com
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