[Histonet] Osmium tetroxide staining for lipids

[Houston, Ronald]

Barry,

I suspect they are wanting to employ this on paraffin sections as Sheila mentions using it on routine 4 micron sections>



If that is the case, there are published articles using Sudan Black B and Oil Red O (Histopathology 2002, 41, 75-79) in paraffin sections, and I believe Tim Morken has used OsO4 in paraffin sections.



I have heard of fixed blocks being impregnated with Osmium before processing and then being sectioned; the Osmium bound lipids are stained.



I've even used en-bloc Osmium staining for degenerating myelin with the Marchi technique (now there's a walk down memory lane!!)



All that being said, I would not recommend using OsO4 in a lab that has not previously used it, as it is an extremely hazardous substance to use, and there are other alternatives.



As a matter of interest Sheila, did your pathologist mention what he was wanting to specifically use the osmium for, or is this just some stab in the dark because he read it in some obscure journal?



Ronnie Houston, MS HT(ASCP)QIHC
Anatomic Pathology Manager
ChildLab, a Division of Nationwide Children's Hospital
www.childlab.com

700 Children's Drive
Columbus, OH 43205
(P) 614-722-5450
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www.NationwideChildrens.org<http://www.nationwidechildrens.org/>

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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Rittman, Barry R
Sent: Friday, August 24, 2012 9:08 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Osmium tetroxide staining for lipids



Sheila Hi

I am assuming that you are using frozen sections?

The easiest way is to put a small amount of aqueous osmium tetroxide in the bottom of a dish that has a tight fitting lid.

Place the slide with section into the dish but not touching the fluid.

Place tightly fitting lid and leave - not sure how long, probably 20- 30 minutes should do it although can leave longer.

Then simply rinse and that's it.

The osmium vapor fixes the lipid so that it is not important to have any buffer in the solution, can just use straightforward osmium tetroxide in distilled water.

The osmium black that forms is insoluble in most solvents.

You should do this in a fume hood as the vapor will fix your nasal mucosa if you are not careful.

Hope that your pathologist realizes that although the osmium will stain the lipid it  makes staining of other components very difficult.

Barry



From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Sheila Adey [sadey <@t> hotmail.ca]

Sent: Friday, August 24, 2012 7:30 AM

To: histonet <@t> lists.utsouthwestern.edu

Subject: [Histonet] Osmium tetroxide staining for lipids



Hi Everyone:



One of my pathologists wants me to look into Osmium Tetroxide for staining lipids. From what I can gather on the internet, it looks like it is used in Electron microscopy for fixation and staining.

Is anyone using this procedure for routine 4 micrometer sections?



Thanks

:)

Sheila                                    _______________________________________________

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