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High Resolution FESEM and TEM Reveal Bacterial Spore Attachment

Published online by Cambridge University Press:  16 July 2007

Barbara J. Panessa-Warren ,
George T. Tortora  and
John B. Warren
Barbara J. Panessa-Warren
Affiliation:
Department of Condensed Matter Physics and Materials Science, Building 480, Brookhaven National Laboratory, Upton, NY 11973, USA
George T. Tortora
Affiliation:
Clinical Microbiology Laboratory, University Hospital, SUNY Stony Brook, Stony Brook, NY 11794, USA School of Health Technology & Management, Stony Brook University, Stony Brook, NY 11794, USA
John B. Warren
Affiliation:
Division of Instrumentation, Building 535B, Brookhaven National Laboratory, Upton, NY 11973, USA
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Abstract

Transmission electron microscopy (TEM) studies in the 1960s and early 1970s using conventional thin section and freeze fracture methodologies revealed ultrastructural bacterial spore appendages. However, the limited technology at that time necessitated the time-consuming process of imaging serial sections and reconstructing each structure. Consequently, the distribution and function of these appendages and their possible role in colonization or pathogenesis remained unknown. By combining high resolution field emission electron microscopy with TEM images of identical bacterial spore preparations, we have been able to obtain images of intact and sectioned Bacillus and Clostridial spores to clearly visualize the appearance, distribution, resistance (to trypsin, chloramphenicol, and heat), and participation of these structures to facilitate attachment of the spores to glass, agar, and human cell substrates. Current user-friendly commercial field emission scanning electron microscopes (FESEMs), permit high resolution imaging, with high brightness guns at lower accelerating voltages for beam sensitive intact biological samples, providing surface images at TEM magnifications for making direct comparisons. For the first time, attachment structures used by pathogenic, environmental, and thermophile bacterial spores could be readily visualized on intact spores to reveal how specific appendages and outer spore coats participated in spore attachment, colonization, and invasion.

Keywords

field emission scanning electron microscopyClostridium and Bacillus sporesspore attachmenthuman cellstransmission electron microscopy
Type
BIOLOGICAL APPLICATIONS
Information
Microscopy and Microanalysis , Volume 13 , Issue 4 , August 2007 , pp. 251 - 266
Copyright
© 2007 Microscopy Society of America

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