Potential problems with fluorescein diacetate assays of cell viability when testing natural products for antimicrobial activity

Joanne M. Clarke, Michael R. Gillings, Nanda Altavilla, Andrew J. Beattie

Research output: Contribution to journalArticlepeer-review

49 Citations (Scopus)

Abstract

There are two potential problems in the use of fluorescein diacetate (FDA) as a measure of cell viability. The first is the hydrolysis of FDA to fluorescein in the absence of live cells and the second is the quenching of fluorescence by assay solutions. We show that common media components such as tryptone, peptone and yeast extract all promote hydrolysis of FDA in the absence of live cells, as do Tris-HCl and sodium phosphate buffers. As a consequence, various microbiological media promote hydrolysis of FDA in the absence of live cells. Different media were also shown to reduce the amount of visible fluorescence of fluorescein. Diluting the medium decreases the background hydrolysis of FDA as well as increases the amount of visible fluorescence. Both problems should be considered when using FDA as an indicator of cell viability when testing natural products for antimicrobial activity.

Original languageEnglish
Pages (from-to)261-267
Number of pages7
JournalJournal of Microbiological Methods
Volume46
Issue number3
DOIs
Publication statusPublished - 2001

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