Isolation of lipid-rich marine microalgae by flow cytometric screening with Nile Red staining

Katayama, Tsuneo; Kishi, Masatoshi; Takahashi, Kazutaka; Furuya, Ken; Wahid, Mohd Effendy Abd; Khatoon, Helena; Kasan, Nor Azman

doi:10.1007/s10499-019-00344-y

Cited by 16 publications

(6 citation statements)

References 25 publications

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“…BODIPY 505/515 and Nile Red are fluorescent dyes that preferentially stain microalgal cellular lipids ( Govender et al, 2012 ; Sitepu et al, 2012 ; Katayama et al, 2019 ). For many years, BODIPY and Nile Red have been used as microalgal lipid probes and their mechanisms are beginning to be understood.…”

Section: Resultsmentioning

confidence: 99%

Physiological and Biochemical Responses of Bicarbonate Supplementation on Biomass and Lipid Content of Green Algae Scenedesmus sp. BHU1 Isolated From Wastewater for Renewable Biofuel Feedstock

et al. 2022

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In the present study, different microalgae were isolated from wastewater environment and evaluated for higher growth and lipid accumulation. The growth adaptability of all the isolated microalgae were tested for carbon source with supplementation of sodium bicarbonate in BG-11 N+ medium. Further based on the uptake rate of sodium bicarbonate and growth behavior, microalgal strains were selected for biofuel feedstock. During the study, growth parameters of all the isolates were screened after supplementation with various carbon sources, in which strain Scenedesmus sp. BHU1 was found highly effective among all. The efficacy of Scenedesmus sp. BHU1 strain under different sodium bicarbonate (4–20 mM) concentration, in which higher growth 1.4 times greater than control was observed at the concentration 12 mM sodium bicarbonate. In addition, total chlorophyll content (Chl-a + Chl-b), chlorophyll fluorescence (Fv/Fm, Y(II), ETR max, and NPQmax), and biomass productivity were found to be 11.514 μg/ml, 0.673, 0.675, and 31.167 μmol electrons m−2 s−1, 1.399, 59.167 mg/L/day, respectively, at the 12 mM sodium bicarbonate. However, under optimum sodium bicarbonate supplementation, 56.920% carbohydrate and 34.693% lipid content were accumulated, which showed potential of sodium bicarbonate supplementation in renewable biofuel feedstock by using Scenedesmus sp. BHU1 strain.

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Section: Resultsmentioning

confidence: 99%

Physiological and Biochemical Responses of Bicarbonate Supplementation on Biomass and Lipid Content of Green Algae Scenedesmus sp. BHU1 Isolated From Wastewater for Renewable Biofuel Feedstock

et al. 2022

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“…NR has been suggested for fluorescently staining microplastics (Cole, 2016;Shim et al, 2016;Wiggin and Holland, 2019). The use of NR has also been demonstrated in conjunction with FC, wherein microalgae samples were stained by NR in acetone at a concentration of 3 µM and analyzed using flow cytometric cell sorting (Katayama et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

Preliminary Results From Detection of Microplastics in Liquid Samples Using Flow Cytometry

et al. 2020

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Microplastics are globally recognized as contaminants in freshwater and marine aquatic systems. To date there is no universally accepted protocol for isolation and quantification of microplastics from aqueous media. Various methodologies exist, many of which are time consuming and have the potential to introduce contaminants into samples, thereby obscuring characterization of the environmental microplastic load. Here, we present first steps in the detection of microplastics in liquid samples, based on their fluorescent staining followed by high throughput analysis and quantification using Flow Cytometry. Using controlled laboratory settings nine polymer types [polystyrene (PS); polyethylene (PE); polyethylene terephthalate (PET/PETE); high density polyethylene (HDPE); low density polyethylene (LDPE); polyvinyl chloride (PVC); polypropylene (PP); nylon (PA); polycarbonate (PC)] were tested for identification and quantification in freshwater. All nine plastic types were stained with 10 µg/mL Nile Red in 10% dimethyl sulfoxide with a 10 min incubation time. The lowest spatial detectable limit for plastic particles was 200 nm. Out of the nine polymer types chosen for the study PS, PE, PET, and PC were well-identified; however, results for other plastic types (PVC, PP, PA, LDPE, and HDPE) were masked to certain extent by Nile Red aggregation and precipitation. The methodology presented here permits identification of a range of particle sizes and types. It represents a significant step in the quantification of microplastics by replacing visual data interpretation with a sensitive and automated method.

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“…It includes technologies that can automatically count cells, analyze their vitality, size and granularity, and identify multiple physiological states and enzyme activity with a speed reaching thousands of events per second, based on quantified scattered, fluorescent light signals [ 136 , 145 ]. This analysis method can be utilized to isolate and sort desired overproducing mutants [ 146 , 147 ], especially when combined with specific staining dyes, such as Nile Red [ 148 , 149 , 150 ] and BIODIPY [ 149 , 151 ], which are commonly used for intracellular lipid detection to isolate lipid-rich microalgae strains. Despite its numerous advantages, one of its main drawbacks is that extracellular target products cannot be analyzed easily, as their fluorescence signals are not associated with the cells [ 141 , 152 ].…”

Section: Overview Of High-throughput Screening Methods and Techniques...mentioning

confidence: 99%

Strain Development in Microalgal Biotechnology—Random Mutagenesis Techniques

Bleisch

Freitag

Ihadjadene

et al. 2022

Life

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Microalgal biomass and metabolites can be used as a renewable source of nutrition, pharmaceuticals and energy to maintain or improve the quality of human life. Microalgae’s high volumetric productivity and low impact on the environment make them a promising raw material in terms of both ecology and economics. To optimize biotechnological processes with microalgae, improving the productivity and robustness of the cell factories is a major step towards economically viable bioprocesses. This review provides an overview of random mutagenesis techniques that are applied to microalgal cell factories, with a particular focus on physical and chemical mutagens, mutagenesis conditions and mutant characteristics.

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Isolation of lipid-rich marine microalgae by flow cytometric screening with Nile Red staining

Cited by 16 publications

References 25 publications

Physiological and Biochemical Responses of Bicarbonate Supplementation on Biomass and Lipid Content of Green Algae Scenedesmus sp. BHU1 Isolated From Wastewater for Renewable Biofuel Feedstock

Physiological and Biochemical Responses of Bicarbonate Supplementation on Biomass and Lipid Content of Green Algae Scenedesmus sp. BHU1 Isolated From Wastewater for Renewable Biofuel Feedstock

Preliminary Results From Detection of Microplastics in Liquid Samples Using Flow Cytometry

Strain Development in Microalgal Biotechnology—Random Mutagenesis Techniques

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