Quantification of confocal images of biofilms grown on irregular surfaces

Stacy Sommerfeld Ross; Mai Han Tu; Megan L Falsetta; Margaret R Ketterer; Megan R Kiedrowski; Alexander R Horswill; Michael A Apicella; Joseph M Reinhardt; Jennifer Fiegel

doi:10.1016/j.mimet.2014.02.020

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Quantification of confocal images of biofilms grown on irregular surfaces

Journal article

Peer reviewed

Quantification of confocal images of biofilms grown on irregular surfaces

Stacy Sommerfeld Ross, Mai Han Tu, Megan L Falsetta, Margaret R Ketterer, Megan R Kiedrowski, Alexander R Horswill, Michael A Apicella, Joseph M Reinhardt and Jennifer Fiegel

Journal of microbiological methods, Vol.100(1), pp.111-120

05/2014

DOI: 10.1016/j.mimet.2014.02.020

PMCID: PMC4041150

PMID: 24632515

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https://www.ncbi.nlm.nih.gov/pmc/articles/4041150View

Open Access

Abstract

Bacterial biofilms grow on many types of surfaces, including flat surfaces such as glass and metal and irregular surfaces such as rocks, biological tissues and polymers. While laser scanning confocal microscopy can provide high-resolution images of biofilms grown on any surface, quantification of biofilm-associated bacteria is currently limited to bacteria grown on flat surfaces. This can limit researchers studying irregular surfaces to qualitative analysis or quantification of only the total bacteria in an image. In this work, we introduce a new algorithm called modified connected volume filtration (MCVF) to quantify bacteria grown on top of an irregular surface that is fluorescently labeled or reflective. Using the MCVF algorithm, two new quantification parameters are introduced. The modified substratum coverage parameter enables quantification of the connected-biofilm bacteria on top of the surface and on the imaging substratum. The utility of MCVF and the modified substratum coverage parameter were shown with Pseudomonas aeruginosa and Staphylococcus aureus biofilms grown on human airway epithelial cells. A second parameter, the percent association, provides quantified data on the colocalization of the bacteria with a labeled component, including bacteria within a labeled tissue. The utility of quantifying the bacteria associated with the cell cytoplasm was demonstrated with Neisseria gonorrhoeae biofilms grown on cervical epithelial cells. This algorithm provides more flexibility and quantitative ability to researchers studying biofilms grown on a variety of irregular substrata. •Quantify connected-biofilm bacteria grown on visible irregular surfaces•Introduce modified connected volume filtration algorithm•Novel modified substratum coverage parameter assesses bacteria coverage on surface.•Quantify percent bacteria associated with visible surface (fluorescent/reflective)

Biofilm

COMSTAT

Confocal microscopy

Quantification

Surface

Tissue

Details

Title: Subtitle: Quantification of confocal images of biofilms grown on irregular surfaces
Creators: Stacy Sommerfeld Ross - University of Iowa
Mai Han Tu - University of Iowa
Megan L Falsetta - University of Iowa
Margaret R Ketterer - University of Iowa
Megan R Kiedrowski - University of Iowa
Alexander R Horswill - University of Iowa
Michael A Apicella - University of Iowa
Joseph M Reinhardt - University of Iowa
Jennifer Fiegel - University of Iowa
Resource Type: Journal article
Publication Details: Journal of microbiological methods, Vol.100(1), pp.111-120
DOI: 10.1016/j.mimet.2014.02.020
PMID: 24632515
PMCID: PMC4041150
NLM abbreviation: J Microbiol Methods
ISSN: 0167-7012
eISSN: 1872-8359
Publisher: Elsevier B.V
Grant note: DOI: 10.13039/100000060, name: National Institute of Allergy and Infectious Diseases, award: AI096007, AI078921; DOI: 10.13039/100001797, name: PhRMA; DOI: 10.13039/100000002, name: NIH, award: UL1RR024979
Language: English
Date published: 05/2014
Academic Unit: Roy J. Carver Department of Biomedical Engineering; Radiology; Microbiology and Immunology; Pharmaceutical Sciences and Experimental Therapeutics; Chemical and Biochemical Engineering; Internal Medicine
Record Identifier: 9984196987102771

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