By John J. Bozzola
Electron Microscopy Covers all the very important points Of Electron Microscopy For Biologists, together with concept Of Scanning And Transmission, Specimen coaching, electronic Imaging And picture research, Laboratory security And Interpretation of pictures. The textual content additionally encompasses a entire Atlas Of Ultrastructure.
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Additional info for Electron Microscopy, 2nd Edition
Calcium or magnesium chloride is often added to the buffer to preserve certain features within the tissue (such as membranes and DNA). They become airborne and are accidentally inhaled. Page 24 Obtaining and Preparing Buffered Glutaraldehyde Fixative Assuming a standard glutaraldehyde prefixation and osmium tetroxide postfixation combination is to be used, the preparation of a solution of 3% buffered glutaraldehyde and another of 1% osmium tetroxide will be described. Biological grade glutaraldehyde is less pure, but is considered adequate for most perfusion fixation purposes.
This may be accomplished using modified cryo pliers, precooled silver hammers, or, most efficiently, using commercially available devices that gravitydrop a rod containing the specimen mounted on the end onto the chilled metal. Vitrification is achieved to a depth of 10 to 15 m. The rapidly flowing jet of propane achieves vitrification of the specimen to a total depth of approximately 40 m. To retrieve the sample, the two metal plates are separated under liquid nitrogen and the specimen further processed as desired.
Glutaraldehyde can be expected to penetrate at a rate of less than 1 mm per hour if compact tissue is immersed into the glutaraldehyde solution. Therefore, to obtain rapid immersion fixation, the immersed tissue should be under 1 mm in thickness in at least one dimension. This is especially well demonstrated when tissues are perfused with glutaraldehyde. Glutaraldehyde should be used under a wellventilated hood and disposed of in a proper manner. The osmium tetroxide molecule is symmetrical and contains four doublebonded oxygen molecules.
Electron Microscopy, 2nd Edition by John J. Bozzola