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Measuring the thickness of an outer layer of viable bacteria in an oral biofilm by viability mapping

Hope, C.K. and Wilson, M. (2003) Measuring the thickness of an outer layer of viable bacteria in an oral biofilm by viability mapping. Journal of Microbiological Methods, 54 (3). pp. 403-410. ISSN 0167-7012

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Abstract

Bacterial biofilms have been reported to contain distinct regions of viable and nonviable bacteria. The purpose of this study was to identify such regions in biofilms of oral bacteria and to determine their dimensions. Oral biofilms were grown aerobically in a constant-depth film fermenter and studied using confocal laser scanning microscopy (CLSM) incorporating viability staining with water immersion lenses. A variety of viability distributions were observed, including biofilm “stacks” possessing an outer layer of viable bacteria surrounding an internal core of nonviable bacteria. Using image analysis tools, we measured the thickness of this outer viable region, in the x-y plane, from single confocal optical sections, and determined the mean angle () of these portions of the biofilm stack (10.93º). x-y plane thickness data in conjunction with the data on the angle of the stack returned the thickness of the outer viable layer perpendicular to the bulk medium flow as 36.62 μm (31.61 – 42.21 μm accounting for 95% confidence for variation in both the x-y plane thickness and θ). We have shown that CLSM, in conjunction with vital stains and image analysis techniques, can reveal viability patterns in biofilms and where appropriate can be used to measure the dimensions of these structures.

Item Type:Article
Additional Information:Issue: September 2003. Available online 1 May 2003.
Uncontrolled Keywords:Biofilm; Viability mapping; Confocal laser scanning microscopy; Viable staining; ImageJ ; MICROCOSM DENTAL PLAQUES; IMAGE THRESHOLDING METHODS; LASER-SCANNING MICROSCOPY; SPATIAL-DISTRIBUTION; FILM FERMENTER; CHLORHEXIDINE; SUSCEPTIBILITY; GROOVES
Subjects:R Medicine > RK Dentistry
Q Science > QR Microbiology
Departments, Research Centres and Related Units:Academic Faculties, Institutes and Research Centres > Faculty of Medicine > School of Dental Sciences
DOI:10.1016/S0167-7012(03)00085-X
Refereed:Yes
Status:Published
ID Code:66
Deposited On:11 Apr 2008 16:04
Last Modified:20 Jan 2012 14:43

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