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Correlating Fluorescence Microscopy with Electron Microscopy

Published online by Cambridge University Press:  14 March 2018

Jon Charlesworth
Affiliation:
Mayo Clinic

Extract

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The use of fluorescent probes is becoming more and more common in cell biology. It would be useful if we were able to correlate a fluorescent structure with an electron microscopic image. The ability to definitively identify a fluorescent organelle would be very valuable. Recently, Ying Ren, Michael Kruhlak, and David Bazett-Jones devised a clever technique to correlate a structure visualized in the light microscope, even a fluorescing cell, with transmission electron microscopy (TEM).

Two keys to the technique of Ren et al are the use of grids (as used in the TEM) with widely spaced grid bars and the use of Quetol as the embedding resin. The grids allow for cells to be identified between the grid bars, and in turn the bars are used to keep the cell of interest in register throughout the processing for TEM. Quetol resin was used for embedding because of its low auto fluorescence and sectioning properties. The resin also becomes soft and can be cut and easily peeled from glass coverslips when heated to 70°C.

Type
Research Article
Copyright
Copyright © Microscopy Society of America 2004

Footnotes

1

The authors gratefully acknowledge Dr. David Brazett-Jones for reviewing this article, Jon Charlesworth is the Coordinator of die Electron Microscopy Core Facility at Mayo Clinic.

References

Note

2. Ren, Y., M.J. Kruhlak, and D.P. Bazett-Jones, Same serial section correlative light and energy-filtered transmission electron microscopy, J. Histochem. Cytochem. 51:605-612, 2003.