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Published (Version of record)CC BY V4.0, Open Access
Published (Version of record)CC BY V4.0, Open
Journal article
Colloid chemistry pitfall for flow cytometric enumeration of viruses in water
Water research X, Vol.2, 100025
01/02/2019
PMCID: PMC6549941
PMID: 31194069
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Source: InCites
Abstract
Flow cytomtery (FCM) has become a standard approach to enumerate viruses in water research. However, the nature of the fluorescent signal in flow cytometric analysis of water samples and the mechanism of its formation, have not been addressed for bacteriophages expected in wastewaters. Here we assess the behaviour of fluorescent DNA-staining dyes in aqueous solutions, as well as sensitivity and accuracy of FCM for enumeration of DNA-stained model bacteriophages λ, P1, and T4. We demonstrate that in aqueous systems fluorescent dyes form a self-stabilized (pseudolyophilic) emulsion of auto-fluorescing colloid particles. Sample shaking and addition of surfactants enhance auto-fluorescence due to increased dispersion and, in the presence of surfactants, stabilization of the dye emulsion. Bacteriophages with genome sizes <100 kbp (i.e. λ & P1) did not generate a distinct population signal to be detected by one of the most sensitive FCM instruments available (BD LSR Fortessa™ X-20), whereas the larger T4 bacteriophage was resolved as a distinct population of events. These results indicate that the use of fluorescent dyes for bacteriophage enumeration by flow cytometry can produce false positive signals and lead to wrong estimation of total virus counts by misreporting colloid particles as virions, depending on instrument sensitivity.
Details
- Title
- Colloid chemistry pitfall for flow cytometric enumeration of viruses in water
- Creators
- Elena A. Dlusskaya - University of AlbertaAlexey M. Atrazhev - University of AlbertaNicholas J. Ashbolt - University of Alberta
- Publication Details
- Water research X, Vol.2, 100025
- Publisher
- Elsevier
- Identifiers
- 991013098098302368
- Copyright
- © 2019 The Author(s). Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
- Academic Unit
- Faculty of Science and Engineering
- Language
- English
- Resource Type
- Journal article