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Localization of Green Fluorescent Protein Absorbance by Energy Filtered Transmission Electron Microscopy

Published online by Cambridge University Press:  02 July 2020

J. A. Davis ,
R. G. Garces ,
J.-Y. Diao  and
F. P. Ottensmeyer
J. A. Davis
Affiliation:
Department of Medical Biophysics, University of Toronto, Toronto, ONM5G 2M9
R. G. Garces
Affiliation:
Department of Medical Biophysics, University of Toronto, Toronto, ONM5G 2M9
J.-Y. Diao
Affiliation:
Department of Medical Biophysics, University of Toronto, Toronto, ONM5G 2M9
F. P. Ottensmeyer
Affiliation:
Department of Medical Biophysics, University of Toronto, Toronto, ONM5G 2M9
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Extract

Energy filtered transmission electron microscopy has the potential to provide high resolution, spatially resolved, atomic and chemical information. However, aberrations generated by the electron spectrometer blur the energy resolution and limit the atomic or molecular distributions that can be studied. Energy absorptions corresponding to the visible light range fall below an energy loss of 5 eV. The selection of electrons that have lost an amount of energy corresponding to chromophore absorption by the sample thus requires a spectrometer with a high energy resolution over the full image plane. A corrected prism-mirror-prism filter that has a resolution of 1.1 eV, sufficient to select these low energy loss electrons, was developed and installed by us in a Zeiss EM902. Its imaging capability was verified for a number of different chromophores. The chromophore currently under study is that of the green fluorescent protein (GFP).

Type
Labeling for Microscopy and Correlative Microscopy
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. 324 - 325
Copyright
Copyright © Microscopy Society of America

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References

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