Title

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

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 μl 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.

Publication Date

1-1-1997

Publication Title

Biophysical Journal

Volume

72

Issue

4

Number of Pages

1521-1526

Document Type

Article

Personal Identifier

scopus

DOI Link

https://doi.org/10.1016/S0006-3495(97)78800-3

Socpus ID

0030901113 (Scopus)

Source API URL

https://api.elsevier.com/content/abstract/scopus_id/0030901113

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