Please use this identifier to cite or link to this item: https://hdl.handle.net/1822/35134

TitleQuantification and viability analyses of Pseudokirchneriella subcapitata algal cells using image-based cytometry
Author(s)Machado, Manuela D.
Soares, Eduardo V.
KeywordsAutomated algae counting
Copper stress
Fluorescent dye
Image-based cytometry
Quantitative measure of algae viability
SYTOX Green
Issue dateApr-2015
PublisherSpringer
JournalJournal of Applied Phycology
CitationMachado, Manuela D.; Soares, Eduardo V., Quantification and viability analyses of Pseudokirchneriella subcapitata algal cells using image-based cytometry. Journal of Applied Phycology, 27(2), 703-710, 2015
Abstract(s)This work aims to evaluate the feasibility of using image-based cytometry (IBC) in the analysis of algal cell quantification and viability, using Pseudokirchneriella subcapitata as a cell model. Cell concentration was determined by IBC to be in a linear range between 1×105 and 8×106 cells mL1. Algal viability was defined on the basis that the intact membrane of viable cells excludes the SYTOX Green (SG) probe. The disruption of membrane integrity represents irreversible damage and consequently results in cell death. Using IBC, we were able to successfully discriminate between live (SG-negative cells) and dead algal cells (heat-treated at 65 °C for 60 min; SG-positive cells). The observed viability of algal populations containing different proportions of killed cells was well correlated (R 2=0.994) with the theoretical viability. The validation of the use of this technology was carried out by exposing algal cells of P. subcapitata to a copper stress test for 96 h. IBC allowed us to follow the evolution of cell concentration and the viability of copper-exposed algal populations. This technology overcomes several main drawbacks usually associated with microscopy counting, such as labour-intensive experiments, tedious work and lack of the representativeness of the cell counting. In conclusion, IBC allowed a fast and automated determination of the total number of algal cells and allowed us to analyse viability. This technology can provide a useful tool for a wide variety of fields that utilise microalgae, such as the aquatic toxicology and biotechnology fields.
TypeArticle
URIhttps://hdl.handle.net/1822/35134
DOI10.1007/s10811-014-0377-4
ISSN0921-8971
1573-5176
e-ISSN1573-5176
Publisher versionhttp://link.springer.com/article/10.1007%2Fs10811-014-0377-4
Peer-Reviewedyes
AccessOpen access
Appears in Collections:CEB - Publicações em Revistas/Séries Internacionais / Publications in International Journals/Series

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