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Unit Code 4993:
Electron Microscopy

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Method Description

The fixed tissues received are rinsed three times in 0.1 M sodium

phosphate buffer, post-fixed, and stained in 2% osmium tetroxide.

The tissues are then rinsed 3 times in distilled water, en bloc

stained in 2% aqueous uranyl acetate (60 degrees C) for 30 minutes,

and dehydrated in a graded series of ethanol and propylene oxide.  

The tissues are subsequently infiltrated and embedded in Spurr's

low viscosity embedding resin. Semi-thin (1 micron) sections for light

microscopy are cut with a Reichert-Jung Ultramicrotome and stained

with toluidine blue. Thin sections are cut with a Reichert-Jung

Ultramicrotome, post-stained with 0.3% aqueous lead citrate, and

examined with a Philips CM12/STEM or JEOL 1200 EXII transmission

electron microscope operated at an appropriate kV. Exposures are

made at 1,000 - 500,000 magnification on Kodak 4489 electron

microscope film. Finished 8 x 10 electron micrograph prints are

produced at 2.5 enlargement factor on Kodak paper. (Spurr AR:  

A low-viscosity epoxy resin embedding medium for electron

microscopy. J Ultrastuc Res 1969;26:31-43; McDowell EM, Trump BF:

Histologic fixatives suitable for diagnostic light and electron

microscopy. Arch Pathol Lab Med 1976;100:405-414)

Performing Laboratory Location

Rochester

Key