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Microwave Processing of Drosophila Tissues for Electron Microscopy

Published online by Cambridge University Press:  14 March 2018

JoAnn Buchanan
JoAnn Buchanan*
Affiliation:
Stanford University School of Medicine
*
redhair@stanford.edu

Extract

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Insect tissue is often difficult to prepare for electron microscopy because of the impenetrable barrier surrounding the body tissues. Drosophila salivary glands have been used for numerous studies because of the large size of the cells and their large polytene chromosomes. Early TEM studies of salivary glands used a protocol that took several days. We were able to achieve excellent preservation and good ultrastructure in Drosophila salivary glands and imaginal discs from Stage L3 larvae using microwave processing in a protocol requiring less than 2 hours.

We used a Pelco Laboratory microwave (model #3451) equipped with a Cold Spot, Steadytemp chiller/recirculator run at 15° C, and vacuum chamber (Ted Pella, Mountain Lakes, CA). The heads and attached salivary glands were removed from the animals and placed in PBS. The tissue was transferred to Pelco prep-eze specimen holders (#36157) for ease of handling. Our goal was to use the microwave effect, not the heating effect, to prepare the tissue.

Type
Microscopy 101
Information
Microscopy Today , Volume 12 , Issue 6 , November 2004 , pp. 42
Copyright
Copyright © Microscopy Society of America 2004

References

1. Lane, N.J. and Y.R., Carter. Puffs and Salivary Gland Function: The Fine Structure of the Larval and Prepupal Salivary Glands of Drosophila melanogaster. Wilhelm Roux’ Archive 169:216–238, 1972.Google Scholar
2. Giberson, R.T. Vacuum-Assisted Microwave Processing of Animal Tissues for Electron Microscopy in Microwave Techniques and Protocols. Giberson, R.T. and Demaree, R.S., pp. 13–23, Humana press, Totowa, New Jersey, 2001.Google Scholar
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