Rapid detection of neutral lipid in green microalgae by flow cytometry in combination with Nile red staining—an improved technique

Satpati, Gour Gopal; Pal, Ruma

doi:10.1007/s13213-014-0937-5

Cited by 42 publications

(28 citation statements)

References 62 publications

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“…Although qualitative analysis suggested more LD in the EBC from the stems of control plants compared with salt‐treated plants, a quantitative analysis is required to support this. Isolated EBC taken from the stem (100 cells per sample) were stained with nile red (9‐diethylamino‐5H‐benzo [α]phenoxazine‐5‐one), a lipid‐soluble dye, highly fluorescent in nonpolar hydrophobic environments, and an excellent probe for the detection of neutral lipids (Satpati & Pal, ). Indirect quantification of nile red fluorescence in EBC by flow cytometry showed that the occurrence of LD in discrete populations in EBC from untreated plants compared with salt treated plants was greater, as observed from representative dot plots and flow histograms (Figure a,b).…”

Section: Resultsmentioning

confidence: 99%

“…In contrast, significant differences could be observed in LD in the cytoplasm of EBC from the stem. Here, many more and often bigger LD could be detected in control tissue compared with salt-treated tissue and an excellent probe for the detection of neutral lipids (Satpati & Pal, 2015). Indirect quantification of nile red fluorescence in EBC by (Figure 5a,b).…”

Section: Lipid Droplets Are Present In the Ebc And Show Salinity-inmentioning

confidence: 90%

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Single cell‐type analysis of cellular lipid remodelling in response to salinity in the epidermal bladder cells of the model halophyte Mesembryanthemum crystallinum

Barkla

Garibay-Hernández

Melzer

et al. 2018

Plant Cell & Environment

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Salt stress causes dramatic changes in the organization and dynamic properties of membranes, however, little is known about the underlying mechanisms involved. Modified trichomes, known as epidermal bladder cells (EBC), on the leaves and stems of the halophyte Mesembryanthemum crystallinum can be successfully exploited as a single-cell-type system to investigate salt-induced changes to cellular lipid composition. In this study, alterations in key molecular species from different lipid classes highlighted an increase in phospholipid species, particularly those from phosphatidylcholine and phosphatidic acid (PA), where the latter is central to the synthesis of membrane lipids. Triacylglycerol (TG) species decreased during salinity, while there was little change in plastidic galactolipids. EBC transcriptomic and proteomic data mining revealed changes in genes and proteins involved in lipid metabolism and the upregulation of transcripts for PIPKIB, PI5PII, PIPKIII, and phospholipase D delta suggested the induction of signalling processes mediated by phosphoinositides and PA. TEM and flow cytometry showed the dynamic nature of lipid droplets in these cells under salt stress. Altogether, this work indicates that the metabolism of TG might play an important role in EBC response to salinity as either an energy reserve for sodium accumulation and/or driving membrane biosynthesis for EBC expansion.

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Section: Resultsmentioning

confidence: 99%

Section: Lipid Droplets Are Present In the Ebc And Show Salinity-inmentioning

confidence: 90%

Single cell‐type analysis of cellular lipid remodelling in response to salinity in the epidermal bladder cells of the model halophyte Mesembryanthemum crystallinum

Barkla

Garibay-Hernández

Melzer

et al. 2018

Plant Cell & Environment

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“…After washing, the biomass was transferred to preweighed vials for oven drying at 70 ∘ C to get a constant dry weight. Biomass yield was calculated by the gravimetric method and expressed as g/L [27,34].…”

Section: Measurement Of Dry Cell Weight (Dcw)mentioning

confidence: 99%

“…Fluorescence activated cell sorting (FACS) also can be employed to screen the microalgae from a mixture of cell populations due to the autofluorescence properties of algal native pigments like chlorophylls, carotenoids, and phycobilins [21][22][23][24]. Flow cytometry in combination with a lipophilic fluorescent probe was applied to screen high lipid containing microalgae for several times [25][26][27]. The application of flow cytometry to detect the neutral lipid in stress induced microalgal cells was studied by a few group of researchers.…”

Section: Introductionmentioning

confidence: 99%

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Efficacy of EDTA and Phosphorous on Biomass Yield and Total Lipid Accumulation in Two Green Microalgae with Special Emphasis on Neutral Lipid Detection by Flow Cytometry

Satpati

Gorain

Pal

2016

Advances in Biology

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Chlorella ellipsoidea and Chlorococcum infusionum, promising microalgae for biodiesel feedstock production, were treated with ethylenediaminetetraacetate (EDTA) and phosphorous to induce stress which was then followed by flow cytometry to study the enhanced intracellular neutral lipid content. Treatment resulted in up to a threefold increase in total lipid content of Chlorella (41.8±1.9% at 16 days of incubation period) and more than twofold increases in Chlorococcum (31.3±1.0% at 18 days of incubation period) under phosphorous starvation in the culture. It was observed that maximum biomass yields in Chlorella and Chlorococcum were 1.56±0.06 and 2.17±0.12 g/L at 1.5 g/L of phosphorous after 20 and 18 days of incubation periods, respectively. The qualitative analyses of neutral lipid bodies under stress conditions were performed by confocal microscopy and revealed bright golden-yellow lipid droplets in stress exposed cells. Significant increase of monounsaturated fatty acids under the nutrient limited conditions was suitable to produce biodiesel. The maximum biomass (g/L) and lipid content (% dry cell weight) at different stresses showed significant results (p<0.05) by single-factor Analysis of Variance (ANOVA) followed by Duncan’s Multiple Range Test (DMRT).

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Integration of Microalgae‐Based Wastewater Bioremediation–Biorefinery Process to Promote Circular Bioeconomy and Sustainability: A Review

Singh

Shourie

Mazahar

2022

CLEAN Soil Air Water

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Bioremediation of wastewater using microalgae is inexpensive, energy efficient, and effective in pollutant reduction as compared to conventional wastewater treatment technologies. Wastewater is a huge resource of minerals, nutrients, bioenergy, and valuable organic compounds and can be used for cultivation of microalgae. The microalgal biomass can be further used as biorefinery feedstock to produce biofuels and commercially important high‐value products. The potential of microalgae toward bioremediation and biorefinery applications presents the avenues for integrating the two processes to support circular bioeconomy and sustainability. This review presents a holistic view of integration of bioremediation and biorefinery processes using microalgae for deriving multiple benefits like pollutant removal, resource recovery, biofuel production, and generation of high‐value commercial products. The current status of high‐throughput microalgal screening technologies is also discussed since the selection of suitable microalgal strains is crucial for the application. The review further summarizes various processes involved in bioremediation and biorefinery systems such as cultivation, bioremediation, harvesting, and downstream processing. Recent trends in microalgal strain improvement for bioremediation and biorefinery applications through genetic engineering, bioinformatics, omics technologies, and genome editing tools are highlighted, while addressing the risks, biosafety issues, and regulatory affairs associated with genetically modified algae.

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Rapid detection of neutral lipid in green microalgae by flow cytometry in combination with Nile red staining—an improved technique

Cited by 42 publications

References 62 publications

Single cell‐type analysis of cellular lipid remodelling in response to salinity in the epidermal bladder cells of the model halophyte Mesembryanthemum crystallinum

Single cell‐type analysis of cellular lipid remodelling in response to salinity in the epidermal bladder cells of the model halophyte Mesembryanthemum crystallinum

Efficacy of EDTA and Phosphorous on Biomass Yield and Total Lipid Accumulation in Two Green Microalgae with Special Emphasis on Neutral Lipid Detection by Flow Cytometry

Integration of Microalgae‐Based Wastewater Bioremediation–Biorefinery Process to Promote Circular Bioeconomy and Sustainability: A Review

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