X-ray micrography and imaging of Escherichia coli cell shape using laser plasma pulsed point X-ray sources

    Authors

    J. M. Rajyaguru; M. Kado; M. C. Richardson;M. J. Muszynski

    Comments

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    Abbreviated Journal Title

    Biophys. J.

    Keywords

    ATOMIC-FORCE MICROSCOPY; DNA; RESOLUTION; COMPLEXES; MOLECULES; SURFACE; Biophysics

    Abstract

    High-resolution x-ray microscopy is a relatively new technique and is performed mostly at a few large synchrotron x-ray sources that use exposure times of seconds, We utilized a bench-top source of single-shot laser (ns) plasma to generate x-rays similar to synchrotron facilities, A 5 mu l suspension of Escherichia coli ATCC 25922 in 0.9% phosphate buffered saline was placed on polymethylmethyacrylate coated photoresist, covered with a thin (100 nm) SiN window and positioned in a vacuum chamber close to the x-ray source, The emission spectrum was tuned for optimal absorption by carbon-rich material, Atomic force microscope scans provided a surface and topographical image of differential x-ray absorption corresponding to specimen properties, By using this technique we observed a distinct layer around whole cells, possibly representing the Gram-negative envelope, darker stained areas inside the cell corresponding to chromosomal DNA as seen by thin section electron microscopy, and dent(s) midway through one cell, and 1/3- and 2/3-lengths in another cell, possibly representing one or more division septa, This quick and high resolution with depth-of-field microscopy technique is unmatched to image live hydrated ultrastructure, and has much potential for application in the study of fragile biological specimens.

    Journal Title

    Biophysical Journal

    Volume

    72

    Issue/Number

    4

    Publication Date

    1-1-1997

    Document Type

    Article; Proceedings Paper

    Language

    English

    First Page

    1521

    Last Page

    1526

    WOS Identifier

    WOS:A1997WP36900005

    ISSN

    0006-3495

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