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Strategies For Combining Elemental Distribution Data Derived From Multiple Images and Samples

Published online by Cambridge University Press:  02 July 2020

Marie E. Cantino  and
Joseph G. Eichen
Marie E. Cantino
Affiliation:
Department of Physiology and Neurobiology, University of Connecticut, Storrs, Connecticut, 06269
Joseph G. Eichen
Affiliation:
Sloan-Kettering Research Center, New York, N.Y., 10024
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Data from digital electron probe x-ray microanalysis (EPXMA) images can be used to generate high resolution profiles of Ca binding within sarcomeres of vertebrate striated muscle. While ratios of elemental peak to bremstrahlung at each point (Hall method) have been used in many studies to allow comparison of data taken from different samples, this procedure has limitations for our application. A different normalization procedure is described here which provides a means of assessing variation among elemental and bremstrahlung profiles obtained from different images and samples.

Freeze dried cryosections of chemically skinned frog or rabbit skeletal muscle were prepared as previously described. Digital EPXMA images were collected for 24 to 36 hours using a Zeiss EM910 STEM and an Oxford ExL2 microanalysis system. Calcium and bremstrahlung counts were summed within one pixel wide masks placed at successive positions along each half sarcomere, as previously described, using an automated routine in IPLAB on a Power Mac 7100.

Type
Compositional Mapping With High Spatial Resolution
Information
Microscopy and Microanalysis , Volume 4 , Issue S2: Proceedings: Microscopy & Microanalysis '98, Microscopy Society of America 56th Annual Meeting, Microbeam Analysis Society 32nd Annual Meeting, Atlanta, Georgia July 12-16, 1998 , July 1998 , pp. 134 - 135
Copyright
Copyright © Microscopy Society of America

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References

1. Cantino, M. E., Allen, T. S. and Gordon, A. M. (1993). Biophys. J. 64: 211–222.CrossRefGoogle Scholar

2. Cantino, M. E. and Eichen, J. G.. (1996) Proceedings, Microscopy and Microanalysis 1996 San Francisco Press, Inc. pp. 292–293Google Scholar

3. Hall, T. A. in Oster, G., Ed., Physical Techniques in Biological Research. New York, Academic Press (1971) 12.Google Scholar

4. This research was supported by a grant from the NIH (HL49443).Google Scholar