Microalgal cytometric analysis in the presence of endogenous autofluorescent pigments

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dc.contributor.authorDashkova, Veronika
dc.contributor.authorSegev, Einat
dc.contributor.authorMalashenkov, Dmitry
dc.contributor.authorKolter, Roberto
dc.contributor.authorVorobjev, Ivan
dc.contributor.authorBarteneva, Natasha S.
dc.creatorVeronika, Dashkova
dc.date.accessioned2017-12-22T09:28:34Z
dc.date.available2017-12-22T09:28:34Z
dc.date.issued2016-11-01
dc.description.abstractAbstract Flow cytometry (FCM) is a well-established tool in the field of aquatic phytoplankton ecology and microalgal biotechnology, which allows for rapid assessment of the viability and physiological state of individual cells in algal populations. However, the autofluorescent spectra of different types of chlorophyll and other algal pigments may overlap with fluorescent dyes and affect the resolution of algae clusters, sensitivity, and signal-to-noise ratio. Dying algal cells continue to exhibit a strong autofluorescent signal, which may affect the evaluation of algal viability.Herein, we tested two different approaches to measure algal fluorescence in the presence of a strong autofluorescent signal: 1) by separating dyes between different excitation lasers in order to reach minimal spectral overlap with the autofluorescent signal using flow and imaging cytometry and 2) through full spectrum analysis, virtual filtering and spectral unmixing of dye combinations and algal pigments' autofluorescence via spectral flow cytometry. For this purpose, we used viability dyes from the SYTOX family and lipophilic dyes. Among the dyes tested, the SYTOX Blue (SB) dye had minimal overlap with chlorophyll fluorescence and can be combined with autofluorescence assessment and lipophilic dyes (validated with Emiliania huxleyi algal monocultures). Imaging cytometry provided a detailed characterization of algal subpopulations stained with a combination of fluorescent dyes. A spectral flow cytometer allowed us to analyze environmental phytoplankton samples stained with fluorescent dyes in the presence of strong and heterogeneous autofluorescence from intrinsic algal pigments. We concluded that the multi-color staining of algal samples can be achieved in the presence of strong and diverse algal autofluorescence using dyes with minimal spectral overlap, a multi-laser approach (flow and imaging cytometry) and/or virtual filter and spectral flow cytometry instrumentation. This can open a new page for analytical and cell sorting algal applications.en_US
dc.identifierDOI:10.1016/j.algal.2016.05.013
dc.identifier.citationVeronika Dashkova, Einat Segev, Dmitry Malashenkov, Roberto Kolter, Ivan Vorobjev, Natasha S. Barteneva, Microalgal cytometric analysis in the presence of endogenous autofluorescent pigments, In Algal Research, Volume 19, 2016, Pages 370-380en_US
dc.identifier.issn22119264
dc.identifier.urihttps://www.sciencedirect.com/science/article/pii/S2211926416301692
dc.identifier.urihttp://nur.nu.edu.kz/handle/123456789/3063
dc.language.isoenen_US
dc.publisherAlgal Researchen_US
dc.relation.ispartofAlgal Research
dc.rights.license© 2016 Elsevier B.V. All rights reserved.
dc.subjectMicroalgaeen_US
dc.subjectAutofluorescenceen_US
dc.subjectEmiliania huxleyien_US
dc.subjectFlow and imaging cytometryen_US
dc.subjectSpectral flow cytometryen_US
dc.subjectViabilityen_US
dc.titleMicroalgal cytometric analysis in the presence of endogenous autofluorescent pigmentsen_US
dc.typeArticleen_US
elsevier.aggregationtypeJournal
elsevier.coverdate2016-11-01
elsevier.coverdisplaydateNovember 2016
elsevier.endingpage380
elsevier.identifier.doi10.1016/j.algal.2016.05.013
elsevier.identifier.eid1-s2.0-S2211926416301692
elsevier.identifier.piiS2211-9264(16)30169-2
elsevier.identifier.scopusid85027957244
elsevier.openaccess0
elsevier.openaccessarticlefalse
elsevier.openarchivearticlefalse
elsevier.startingpage370
elsevier.teaserFlow cytometry (FCM) is a well-established tool in the field of aquatic phytoplankton ecology and microalgal biotechnology, which allows for rapid assessment of the viability and physiological state...
elsevier.volume19
workflow.import.sourcescience

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