| From: | Tony Henwood |
Shannon,
Try the following:
Luxol Fast Blue Stain For Myelin
Principle:
Luxol fast blue (Methasol fast blue) is a copper phtalocyanine dye which is employed in myelin demonstration and can be combined with Nissl staining, PAS or van Gieson.
This technique allows easy differentiation of the Luxol Fast Blue.
Fixation: 10% buffered formalin.
Microtomy: Paraffin sections at 5um.
Controls: Spinal Cord
Reagents:
1. Luxol Fast Blue Solution: Warning: Flammable - see MSDS
Luxol fast blue MBS (CI 34200) 0.1g
10% acetic acid 0.5ml
95% ethanol 100ml
Filter. The stain keeps well.
2. 20% ammonia in water.
3. Cresyl Violet counterstain (refer to previous method).
Procedure:
1. Dewax and take sections to alcohol.
2. Stain sections in LFB overnight at 60oC
3. Rinse slides in 95% alcohol followed by distilled water.
4. Differentiate for 10-15 seconds ammonia water. (Until no more blue leaves the section).
5. Wash well in tap water 10-15 minutes.
6. Counterstain in Cresyl Fast Violet 3 minutes.
7. Differentiate in 95% alcohol.
8. Dehydrate, clear and mount.
Results:
Myelin Deep Blue
Nissl Purple
Nuclei Violet
RBCs Pale green
Elastin, collagen Pale green-blue
Reference:
1. Davies R., (1986) "A modified Kluver-Barrera for Myelin and Nissl" Tissue Talk 9(1):10.
Tony Henwood JP, BappSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager
The Children's Hospital at Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: (02) 9845 3306
Fax: (02) 9845 3318
http://www.histosearch.com/homepages/TonyHenwood/default.html
http://us.geocities.com/tonyhenwoodau/index.html
-----Original Message-----
From: schaudoi@mail.mdanderson.org [mailto:schaudoi@mail.mdanderson.org]
Sent: Tuesday, 7 January 2003 1:43
To: histonet@pathology.swmed.edu
Subject: Re: Luxol Fast Blue (Anatomic Stain)
We are looking for a procedure for the Luxol Fast Blue - Anatomic Stain.
If anyone has this procedure could you please send it to me - thanks for your
help in advance.
Shannon Schaffner
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