Photoelectron microscopy and immunofluorescence microscopy of cytoskeletal elements in the same cells
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Date
1983-07
Authors
Nadakavukaren, K. K.
Chen, L. B.
Habliston, D. L.
Griffith, O. H.
Journal Title
Journal ISSN
Volume Title
Publisher
Proceedings of the National Academy of Sciences
Abstract
Pt K2 rat kangaroo epithelial cells and Rat-I fibroblasts
were grown on conductive glass discs, fixed, and permeabilized,
and the cytoskeletal elements actin, keratin , and vimentin
were visualized by indirect immunofluorescence. After the
fluorescence microscopy, the cells were postfixed and dehydrated
for photoelectron microscopy. The contrast in these photoelectron
micrographs is primarily topographical in origin, and the presence
of fluorescent dyes at low density does not contribute significantly
to the material contrast. By comparison with fluorescence
micrographs obtained on the same individual cells, actincontaining
stress fibers, keratin filaments, and vimentin filaments
were identified in the photoelectron micrographs. The apparent
volume occupied by the cytoskeletal network in the cells as judged
from the photoelectron micrographs is much less than it appears
to be from the fluorescence micrographs because the higher resolution
of photoelectron microscopy shows the fibers closer to their
true dimensions. Photoelectron microscopy is a surface technique,
and the images highlight the exposed cytoskeletal structures and
suppress those extending along the substrate below the nuclei. The
results reported here show marked improvement in image quality
of photoelectron micrographs and that this technique has the potential
of contributing to higher resolution studies of cytoskeletal
structures.
Description
5 pages
Keywords
Citation
. Nadakavukaren, K. K., Chen, L. B., Habliston, D. L. & Griffith, O. H. (1983) Photoelectron microscopy and immunofluorescence microscopy of cytoskeletal elements in the same cells. Proc. Natl. Acad. Sci. USA 80, 4012‑4016.