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Updated: Oct 8, 2026

Fluorescence Microscopy Methods for Determining the Viability of Bacteria in Association with Mammalian Cells
Published on: September 5, 2013
The ability of membrane potential dyes and calcafluor white to distinguish between viable and non-viable bacteria
D J Mason1, R Lopéz-Amorós, R Allman
1School of Pure and Applied Biology, University of Wales College of Cardiff, UK.
Abstract:
Various dyes were assessed for their ability to discriminate between viable and non-viable bacteria. Two methods of killing were employed: by heat treatment or by gramicidin treatment. Staining was carried out in two ways; by staining directly in the medium or by washing cells prior to staining in buffer. Carbocyanine and rhodamine 123 dyes only exhibited small changes in fluorescence between viable and non-viable populations of bacteria. Both oxonol dye (bis 1,3-dibutylbarbituric acid trimethine oxonol) and calcafluor white proved much more useful.
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