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Apoptosis Revealed by Epifluorescence can be Compared with Silica Localization by SEM Backscatter using Paraffin Sections on Carbon Planchets

Published online by Cambridge University Press:  02 July 2020

L. Millecchia ,
R. Mercer ,
D. Schwegler-Berry ,
P. Willard  and
S. Friend
L. Millecchia
Affiliation:
Pathology and Physiology Research Branch, Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Morgantown, WV
R. Mercer
Affiliation:
Pathology and Physiology Research Branch, Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Morgantown, WV
D. Schwegler-Berry
Affiliation:
Pathology and Physiology Research Branch, Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Morgantown, WV
P. Willard
Affiliation:
Pathology and Physiology Research Branch, Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Morgantown, WV
S. Friend
Affiliation:
Pathology and Physiology Research Branch, Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Morgantown, WV
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Extract

In investigating pathological changes that occur during development of fibrosis in lungs exposed to silica, we found a significant increase in apoptotic cells in rats exposed to silica by inhalation for 80 days. Since silica is not definitively detected at the light level, we sought to use SEM backscatter imaging to correlate the presence of silica with apoptosis identified by fluorescent assays. To accomplish this, paraffin sections of lungs from rats that had been exposed to silica were placed on carbon planchets. Serial sections were placed on glass slides, immediately before and after the section on the planchet. Using a fluorescein apoptosis detection system (Promega G3250), a TdT-mediated dUTP nick-end labeling (TUNEL) assay was performed on both planchets and slides. The planchets and slides were cover slipped with gel/mount (Biomedia) and viewed in an Olympus AZ70 fluorescent photomicroscope with a wide band blue filter cube. Apoptotic nuclei could be seen in similar regions in both the slide and the planchet preparations.

Type
Pathology
Information
Microscopy and Microanalysis , Volume 6 , Issue S2: Proceedings: Microscopy & Microanalysis 2000, Microscopy Society of America 58th Annual Meeting, Microbeam Analysis Society 34th Annual Meeting, Microscopical Society of Canada/Societe de Microscopie de Canada 27th Annual Meeting, Philadelphia, Pennsylvania August 13-17, 2000 , August 2000 , pp. 578 - 579
Copyright
Copyright © Microscopy Society of America

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References

References:

1.Porter, D.W.,et al, (1999) The Toxicologist 48(1-S): 132.Google Scholar