[Histonet] Electron Microscopy protocols

[Rene J Buesa]

Compressed fat cell cytoplasms (that is what you are seeing as deformed) are cause by to harsh dehydration. Dehydration for TEM specimens has to be extremely graded, without passing from one alcohol to the next with more that 5% gradient.
Additionally fixation has to be complete. Everything in specimens for TEM has to be very gradual, something not very difficult to do in specimens that, as a norm, are 2mm cubes.
Check your processing protocol.
René J.

From: Joel Haas <joel.haas <@t> gladstone.ucsf.edu>
To: histonet <@t> lists.utsouthwestern.edu 
Sent: Wednesday, June 5, 2013 1:29 AM
Subject: [Histonet] Electron Microscopy protocols


Hello all,
    I am working on developing an electron microscopy protocol for looking
at lipid droplets in cultured cells. We have used thin section TEM
previously and found that the morphology of the droplets is often deformed
(should be spherical, shows up as egg shaped or lumpy). If possible we
would like to avoid these types of preservation artifacts. Does anyone have
suggestions for adapting the protocol to better preserve these types of
structures--a triglyceride lipid core surrounded by a phospholipid
monolayer?

Thanks in advance,
Joel
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