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Using confocal laser scanning microscopy, scanning electron microscopy and phase contrast light microscopy to examine marine biofilms

Norton, T.A., Thompson, R.C., Pope, J., Veltkamp, C.J., Banks, B., Howard, C.V. and Hawkins, S.J. (1998) Using confocal laser scanning microscopy, scanning electron microscopy and phase contrast light microscopy to examine marine biofilms Aquatic Microbial Ecology, 16, pp. 199-204. (doi:10.3354/ame).

Record type: Article

Abstract

Confocal laser scanning rnicroscopy (CLSM) was used to examine living organisms within marine biofilms growing on microscope slides and on natural opaque and uneven substrata such as rocks and shells. The results were compared to images of identical fields observed under transmitted llght and phase contrast microscopy and scanning electron microscopy. Confocal microscopy proved superior in sevcrdl respects: (l) It gave clear images of organisms even if they were overlaid with a thick layer of mucus, other organlsms and debr~s. (2) It did not require the biofilm to be disrupted or dned and allowed samples to be re-examined at intervals. (3) It could also distinguish living (fluorescing) organisms from dead cells or inorganic matter. (4) Examining optical 'slices' of samples allowed the 3 dimensional structure of the biofdm to be v~sualised.H owever, species identification, particularly of diatoms, was much easier using scanning electron microscopy. Methods were developed for overlaying a fixed grid on samples so that specific sites or individual organisms could be accurately relocated for re-examination. Appropriate staining methods were also evaluated. Confocal microscopy will prove to be an invaluable aid for examining the structure and growth of living biofilrns in studies of shore ecology and marine fouling.

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More information

Published date: 27 November 1998
Keywords: confocal microscopy, SEM, self-registration, biofilms, microalgae, cyanobacteria, biofouling

Identifiers

Local EPrints ID: 188691
URI: http://eprints.soton.ac.uk/id/eprint/188691
ISSN: 0948-3055
PURE UUID: ea5d1dbc-5881-4b9b-8f8e-35ef19a5a8e3

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Date deposited: 26 May 2011 13:43
Last modified: 18 Jul 2017 11:41

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Contributors

Author: T.A. Norton
Author: R.C. Thompson
Author: J. Pope
Author: C.J. Veltkamp
Author: B. Banks
Author: C.V. Howard
Author: S.J. Hawkins

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