Immunophotoelectron Microscopy: The Electron Optical Analog of Immunofluorescence Microscopy
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Date
1985-01
Authors
Birrell, G. B.
Habliston, D. L.
Nadakavukaren, K. K.
Griffith, O. H.
Journal Title
Journal ISSN
Volume Title
Publisher
Proceedings of the National Academy of Sciences
Abstract
The electron optical analog of immunofluo
rescence microscopy combines three developments: (i) photoelectron
microscopy to produce a high-resolution image of ex
posed components of the cell, (jQ site-specific antibodies, and
(iii) photoemissive markers coupled to the antibodies to make
the distribution of sites visible. This approach, in theory, pro
vides a way to extend the useful immunofluorescence micros
copy technique to problems requiring much higher resolution.
The resolution limit of fluorescence microscopy is limited to
about 200 nm by the wavelength of the light used to form the
image, whereas in photoelectron microscopy the image is
formed by electrons (current resolution: 10-20 nm; theoretical
limit: 5 nm or better depending on the electron optics). As a
test system, cytoskeletons of CV-1 epithelial cells were pre
pared under conditions that preserve microtubules, and the
microtubule networks were visualized by both indirect immu
nofluorescence and immunophotoelectron microscopy using
colloidal gold coated with antibodies. Colloidal gold serves as a
label for immunophotoelectron microscopy, providing en
hanced photoemission from labeled cellular components so
that they stand out against the darker background of the re
maining unlabeled structures. In samples prepared for both
immunofluorescence and immunophotoelectron microscopy,
individual microtubules in the same cells were visualized by
both techniques. The photoemissionof the colloidal gold mark
ers is sufficiently high that the microtubules are easily recog
nized without reference to the immunofluorescence micro
graphs, indicating that this approach can be used, in combina
tion with antibodies, to correlate structure and function in cell
biological studies.
Description
5 pages
Keywords
Citation
Birrell, G. B. , Habliston, D. L., Nadakavukaren, K. K. & Griffith, O. H. (1985) Immunophotoelectron Microscopy: The Electron Optical Analog of Immunofluorescence Microscopy. Proc. Natl. Acad. Sci. USA, 82, 109‑113.