Journal article
Enhanced analysis of bacteria susceptibility in connected biofilms
Journal of microbiological methods, Vol.90(1), pp.9-14
07/2012
DOI: 10.1016/j.mimet.2012.04.008
PMID: 22542520
Abstract
A common method for visualizing bacterial biofilms is through confocal laser scanning microscopy images. Current software packages separate connected-biofilm bacteria from unconnected bacteria, such as planktonic or dispersed bacteria, but do not save both image sequences, making interpretation of the two bacterial populations difficult. Thus we report the development of an algorithm to save separate image sequences and enable qualitative and quantitative evaluation of each bacterial population. To improve bacterial viability assessment using a membrane integrity dye, a colocalization algorithm was also developed. This assigns colocalized pixels to the dead bacteria population, rather than to both the live and dead bacteria groups. Visually, this makes it clearer to distinguish a green live bacteria pixel from a yellow colocalized dead bacteria pixel. This algorithm also aids in the quantification of viability for connected-biofilm bacteria and unconnected bacteria to investigate susceptibility of each population to antimicrobials. The utility of these algorithms was demonstrated with Pseudomonas aeruginosa biofilms treated with ciprofloxacin hydrochloride. Results from this study indicate that quantification with colocalization adjustment can prevent underestimation of dead bacteria. These improvements in image processing will enable researchers to visually differentiate connected-biofilm and unconnected bacteria in a single image and to quantify these populations independently for viability without double counting the colocalized image pixels.
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► Connected-biofilm bacteria separated from unconnected bacteria in confocal images. ► Colocalization adjustment with membrane integrity stain maintained dead bacteria. ► Connected-biofilm bacteria and unconnected bacteria viability quantified separately.
Details
- Title: Subtitle
- Enhanced analysis of bacteria susceptibility in connected biofilms
- Creators
- Stacy Sommerfeld Ross - University of IowaJoseph M Reinhardt - University of IowaJennifer Fiegel - University of Iowa
- Resource Type
- Journal article
- Publication Details
- Journal of microbiological methods, Vol.90(1), pp.9-14
- Publisher
- Elsevier B.V
- DOI
- 10.1016/j.mimet.2012.04.008
- PMID
- 22542520
- ISSN
- 0167-7012
- eISSN
- 1872-8359
- Language
- English
- Date published
- 07/2012
- Academic Unit
- Roy J. Carver Department of Biomedical Engineering; Radiology; Pharmaceutical Sciences and Experimental Therapeutics; Chemical and Biochemical Engineering
- Record Identifier
- 9984197148202771
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