Microalgal cytometric analysis in the presence of endogenous autofluorescent pigments

Dashkova, Veronika; Segev, Einat; Malashenkov, Dmitry; Kolter, Roberto; Vorobjev, Ivan A.; Barteneva, Natasha S.

doi:10.1016/j.algal.2016.05.013

Cited by 23 publications

(9 citation statements)

References 30 publications

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“…However, a spreading spillover from prominent Chl a led to insufficient resolution of different microalgae taxa. The SFC-VF approach [20, 24] allows the creation of “virtual bandpass filters” with no hardware modification and without spectral unmixing. As a result, it was possible to narrow or to widen spectral signal that is taken into consideration from ∼10 nm to ∼300 nm bandwidth (for SP6800 instrument) and to achieve significant discrimination of algal populations.…”

Section: Resultsmentioning

confidence: 99%

Probing complexity of microalgae mixtures with novel spectral flow cytometry approach and “virtual filtering”

Barteneva

Dashkova

Vorobjev

2019

Preprint

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170 27 MS words: 2712 28 29 Abstract 30Fluorescence methods are widely applied for the study of the marine and freshwater 31 phytoplankton communities. However, identification of different microalgae populations by 32 autofluorescent pigments remains a challenge because of the very strong signal from chlorophyll. 33Addressing the issue we developed a novel approach using the flexibility of spectral flow 34 cytometry analysis (SFC) and generated a matrix of virtual filters (VF) capable to of 35 differentiating non-chlorophyll parts of the spectrum. Using this matrix spectral emission regions 36 of algae species were analyzed, and five major algal taxa were discriminated. These results were 37 further applied for tracing particular microalgae taxa in the complex mixtures of laboratory and 38 environmental algal populations. An integrated analysis of single algal events combined with 39 unique spectral emission fingerprints and light scattering parameters of microalgae can be further 40 used to differentiate major microalgal taxa. Our results demonstrate that spectral flow cytometer 41 (SFC-VF) and virtual filtering approach can provide a quantitative assessing of heterogenous 42 phytoplankton communities at single cell level spectra and be helpful in the monitoring of 43 phytoplankton blooms.44 45

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Section: Resultsmentioning

confidence: 99%

Probing complexity of microalgae mixtures with novel spectral flow cytometry approach and “virtual filtering”

Barteneva

Dashkova

Vorobjev

2019

Preprint

Self Cite

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“…Synechocystis sp. PCC 6803 (hereafter called Synechocystis) is an important source of bioproducts that span energy feedstock, cosmetics, and nutraceuticals. , Synechocystis is a spherical, unicellular cyanobacterium with an average cell size of ∼2 μm . Like most bacteria, Synechocystis produces extracellular polymeric substances (EPS) that it uses for aggregation and protection against environmental toxicants .…”

Section: Introductionmentioning

confidence: 99%

“…5,6 Synechocystis is a spherical, unicellular cyanobacterium with an average cell size of ∼2 μm. 7 Like most bacteria, Synechocystis produces extracellular polymeric substances (EPS) that it uses for aggregation and protection against environmental toxicants. 8 EPS, which can comprise as much as 8.4% of the dry weight of Synechocystis, 9 contain carboxyl (≡X-COOH) and phosphoryl (≡X-PO 4 H) groups that are negatively charged at slightly acidic to alkaline conditions.…”

Section: ■ Introductionmentioning

confidence: 99%

Promoting Synechocystis sp. PCC 6803 Harvesting by Cationic Surfactants: Alkyl-Chain Length and Dose Control for the Release of Extracellular Polymeric Substances and Biomass Aggregation

Zhou

Lai

Eustance

et al. 2018

ACS Sustainable Chem. Eng.

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Development of an efficient biomass-harvesting technology for microalgae would achieve cost and energy savings in large-scale microalgae biomass cultivation. Cationic surfactants could improve biomass harvesting, but determining the optimal type and dose of surfactant requires mechanistic understanding. In this study, we evaluated how the alkyl-chain length and dose of three cationic surfactantshexadecyltrimethylammonium bromide (CTAB), myristyltrimethylammonium bromide (MTAB), and dodecyltrimethylammonium bromide (DTAB)affected biomass harvesting of Synechocystis. Flow cytometry (FC) with the nucleic-acid (NA) stain SYTOX Green (SG) was used to differentiate the release of extracellular polymeric substances (EPS) from cell lysis. All selected cationic surfactants dramatically improved the biomass-harvesting efficiency, and harvesting kinetics were represented well with a first-order kinetic model. The efficiency of biomass harvesting correlated positively with the alkyl-chain length; i.e., CTAB > MTAB > DTAB. A longer alkyl chain increased EPS release, which made it easier to achieve a less-negative zeta potential but without cell lysis. For CTAB, the largest cationic surfactant tested, a dose of 4.5 mM and treatment for 60 min achieved the maximum harvesting efficiency of ∼91%. This work lays the foundation for optimizing surfactant species and dose for biomass harvesting.

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“…12, 024111-1 automatic methods of detecting the activity of micro-algae cells have been developed including flow cytometry, Polymerase Chain Reaction (PCR), molecular recognition genomics, and so on. [9][10][11][12][13][14] These methods can determine the activity of micro-algae cells accurately. However, high cost, complex operation, and large volume severely limit the use of these instruments in the on-site detection of micro-algae cells in ship ballast water.…”

Section: Introductionmentioning

confidence: 99%

Detection of viability of micro-algae cells by optofluidic hologram pattern

Wang

et al. 2018

Biomicrofluidics

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A rapid detection of micro-algae activity is critical for analysis of ship ballast water. A new method for detecting micro-algae activity based on lens-free optofluidic holographic imaging is presented in this paper. A compact lens-free optofluidic holographic imaging device was developed. This device is mainly composed of a light source, a small through-hole, a light propagation module, a microfluidic chip, and an image acquisition and processing module. The excited light from the light source passes through a small hole to reach the surface of the micro-algae cells in the microfluidic chip, and a holographic image is formed by the diffraction light of surface of micro-algae cells. The relation between the characteristics in the hologram pattern and the activity of micro-algae cells was investigated by using this device. The characteristics of the hologram pattern were extracted to represent the activity of micro-algae cells. To demonstrate the accuracy of the presented method and device, four species of micro-algae cells were employed as the test samples and the comparison experiments between the alive and dead cells of four species of micro-algae were conducted. The results show that the developed method and device can determine live/dead microalgae cells accurately.

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Microalgal cytometric analysis in the presence of endogenous autofluorescent pigments

Cited by 23 publications

References 30 publications

Probing complexity of microalgae mixtures with novel spectral flow cytometry approach and “virtual filtering”

Probing complexity of microalgae mixtures with novel spectral flow cytometry approach and “virtual filtering”

Promoting Synechocystis sp. PCC 6803 Harvesting by Cationic Surfactants: Alkyl-Chain Length and Dose Control for the Release of Extracellular Polymeric Substances and Biomass Aggregation

Detection of viability of micro-algae cells by optofluidic hologram pattern

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