Colorimetric cell proliferation assay for microorganisms in microtiter plate using water-soluble tetrazolium salts

A colorimetric method to assay cell proliferation of microorganisms in 96-well microtiter plates using water-soluble tetrazolium salts and electron mediators was developed. Combinations of 6 kinds of water-soluble tetrazolium salts and 27 kinds of electron mediators that considered the metabolic eff...

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Bibliographic Details
Published in:Journal of microbiological methods 2008-09, Vol.75 (1), p.109-116
Main Authors: Tsukatani, Tadayuki, Suenaga, Hikaru, Higuchi, Tomoko, Akao, Tetsuyuki, Ishiyama, Munetaka, Ezoe, Kimitoshi, Matsumoto, Kiyoshi
Format: Article
Language:English
Subjects:
Bacteria - growth & development
Bacteria - metabolism
Bacteriological methods and techniques used in bacteriology
Bacteriology
Biological and medical sciences
Cell Proliferation
Cell proliferation assay
Colorimetry - methods
Culture Media - chemistry
Culture Media - metabolism
Electron mediator
Fundamental and applied biological sciences. Psychology
Microbiological Techniques - methods
Microbiology
Microorganism
Quinone
Tetrazolium salt
Tetrazolium Salts - chemistry
Tetrazolium Salts - metabolism
Yeasts - growth & development
Yeasts - metabolism
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Summary:A colorimetric method to assay cell proliferation of microorganisms in 96-well microtiter plates using water-soluble tetrazolium salts and electron mediators was developed. Combinations of 6 kinds of water-soluble tetrazolium salts and 27 kinds of electron mediators that considered the metabolic efficiency of microorganisms and the influence with medium components were investigated. 2-Methyl-1,4-naphthoquinone (NQ) was reduced most effectively by various species of microorganisms, and a combination of WST-8 as a water-soluble tetrazolium salt with 2-methyl-1,4-NQ repressed the increase in background due to medium components. In the presence of 2-methyl-1,4-NQ, WST-8 was reduced by microbial cells to formazan, which exhibited maximum absorbance at 460 nm. The proposed tetrazolium method could be applied to measure proliferations of various microbial cells including 3 kinds of yeast, 9 kinds of Gram-positive bacteria, and 10 kinds of Gram-negative bacteria. Linear relationships between the absorbance and viable microbial cell density were obtained in all microorganisms, suggesting that the absorbance change reflected the microbial cell proliferation.
ISSN:0167-7012
1872-8359
DOI:10.1016/j.mimet.2008.05.016