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Energy Filtering Imaging Of Thick Biological Specimens With In-Column "Omega"-Filter Microscopes.

Published online by Cambridge University Press:  02 July 2020

Mark H. Ellisman
Affiliation:
National Center for Microscopy and Imaging Research, Dept. of Neurosciences, University of California, San Diego, La Jolla, CA, 92093
G.Y. Fan
Affiliation:
National Center for Microscopy and Imaging Research, Dept. of Neurosciences, University of California, San Diego, La Jolla, CA, 92093
T. Honda
Affiliation:
National Center for Microscopy and Imaging Research, Dept. of Neurosciences, University of California, San Diego, La Jolla, CA, 92093
T. Kanayama
Affiliation:
JEOL Ltd., Electron Optics Technical Division, 1-2 Musashino 3-Chome, Akishima, Tokyo 196, Japan
M. Kersker
Affiliation:
JEOL USA, Inc., 11 Dearborn Road, Peabody, MA, 01960
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Extract

Preliminary results were obtained from thick sections of biological specimens using 200 keV electron microscopes with an in-column “Omega” energy filter. Images of selectively stained specimens prepared by osmium impregnation of frog spinal ganglia, with thickness ranging from 0.5 μm to 2.5 μm, show drastic improvement in image quality with the use of the energy filter.

Two instruments with Omega filters were used. One was a non-commercial research prototype at JEOL Ltd. equipped with a LaB6 gun and the other was a commercial instrument (JEM-2010FEF) with a field-emission gun, installed at Kyushu University. We found that, although the field-emission gun offered a brightness 2 to 3 orders of magnitude higher, the LaB6 gun was more suitable for studying thick specimens, primarily due to the higher beam current of the LaB6 gun.

Type
Advances in Instrumentation and Performance
Copyright
Copyright © Microscopy Society of America

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References

1.Lindsey, J.D. and Ellisman, M.H., J. Neuroscience 5 (1985) 3124.CrossRefGoogle Scholar
2. We are grateful to Professor Yoshitsugu Tomokiyo of Kyushu University for allowing us to use his JEM-2010FEF microscope for this study.Google Scholar