[Histonet] Osmium tetroxide staining for lipids

[Smith, Allen]

  Osmium tetroxide solutions will fix your skin on contact, but skin grows back. Osmium tetroxide fumes will fix your corneas which are irreplaceable.
  Osmium tetroxide is dangerous, but it is manageable.  First, make sure your fume hood draws really, really well. Put 100 ml of pH 7.2 phosphate-buffered saline into a 150 or 200 ml glass bottle. Score a 1 gram ampoule of osmium tetroxide with a file. Drop the ampoule into the bottle, close the bottle tightly, and shake it until the ampoule breaks. Let the osmium tetroxide dissolve overnight. Use as little as possible of this solution on frozen sections IN THE FUME HOOD. The glass door should be as far down as is practical. Keep your head out of the fume hood! Only your hands should be inside. Used osmium tetroxide solution and the first rinse should be saved in a tightly closed glass bottle and given to a waste hauler. Since it has salvage value, some waste haulers will take osmium tetroxide without charge. 
   Allen A. Smith, Ph.D.
   Professor of Anatomy 
   Barry University School of Podiatric Medicine
   Miami Shores, Florida

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Friday, August 24, 2012 10:32 AM
To: Sheila Adey; histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Osmium tetroxide staining for lipids

Osmium tetroxide is one of the most dangerous substances you can use in the laboratory. Your pathologist probably read some article or found an old photo of fat "stained" with osmium tetroxide and now wants you to do the same thing.
The problem is that the fat is allowed to react to the fumes of this very nasty substance and this is a very dangerous step.
Nowadays this is never done. If he wants to "demonstrate" fat, freeze the tissue, prepare a frozen section and us Oil Red to demonstrate fat.
This is the most current measure.
On the other hand, as you point out, osmium tetroxide is used in electron microscopy.
René J.


________________________________
From: Sheila Adey <sadey <@t> hotmail.ca>
To: "histonet <@t> lists.utsouthwestern.edu" <histonet <@t> lists.utsouthwestern.edu> 
Sent: Friday, August 24, 2012 8:30 AM
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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