Fluorescence activated cell-sorting principles and applications in microalgal biotechnology

Pereira, Hugo; Schulze, Peter S.C.; Schüler, Lisa; Santos, Tamára; Barreira, Luísa; Varela, João

doi:10.1016/j.algal.2017.12.013

Cited by 68 publications

(63 citation statements)

References 64 publications

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“…Flow cytometry channels with different band‐pass filters can be used for simultaneous detection and distinction of the autofluorescence signals and fluorescent molecules. Overall, by combination of FSC (related to cell size), SSC (related to intracellular granularity), and autofluorescence (related to fluorescent pigments) signals, we can efficiently distinguish different microbial population and sort at single‐cell level …”

Section: Methods Of Flow Cytometry Suitable For Oleaginous Microorganmentioning

confidence: 99%

“…Various fluorescent molecules are available for the specific detection of targeted components within the cell (Table , Table S2). Fluorescent molecules can be categorized based on their mechanism of action . For example, fluorescent dyes Sudan Black B, Nile Red and BODIPY specifically bind with intracellular lipids, whereas Propidium Iodide (PI) specifically binds with nucleic acids.…”

Section: Methods Of Flow Cytometry Suitable For Oleaginous Microorganmentioning

confidence: 99%

“…For example, fluorescent dyes Sudan Black B, Nile Red and BODIPY specifically bind with intracellular lipids, whereas Propidium Iodide (PI) specifically binds with nucleic acids. TOTO‐1, TOTO‐3, SYTOX Green I, SYTOX Green II, SYTO 9, SYTO13, Ethidium Bromide (EtBr), Hoechst 33258/33342, Mithramycin, PicoGreen, Pyronine Y, YOYO‐1, YOPRO‐1 and DAPI were widely used fluorescent molecules for the DNA/RNA detection and quantification as well study of the cell cycles . Specific fluorescent molecules emit higher fluorescence depending on cellular physiological parameters.…”

Section: Methods Of Flow Cytometry Suitable For Oleaginous Microorganmentioning

confidence: 99%

“…Specific fluorescent molecules emit higher fluorescence depending on cellular physiological parameters. For example, cellular pH can be detected using BCECF, and SNARF‐1; membrane potential can be measured using DHR 123, DiBAC, DiOC 6 (3), Oxonol, TMRM and Rhodamine 123 (specific for mitochondrial membrane); enzyme activities can be detected using Carboxyfluoresceindiacetate (CFDA) (related to esterase activity), ELF 97‐phosphate (related to alkaline phosphatase activity), Calcein‐AM, CFSE; Respiratory activity, glucose uptake and reactive oxygen species (ROS) can be measured using CTC, 2‐NBDG, and DHR/DHE, respectively . In most of the microbial bioengineering experiments, FITC has been used to tag proteins to study expression and activities.…”

Section: Methods Of Flow Cytometry Suitable For Oleaginous Microorganmentioning

confidence: 99%

“…In the past few years, research related to oleaginous microalgae, flow cytometry, and their potential applications has increased. A recent review by Pereira et al . detailed FACS principles and applications with special emphasis on microalgal strain development.…”

Section: Applications Of Flow Cytometry In Oleaginous Microorganismsmentioning

confidence: 99%

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Role of flow cytometry for the improvement of bioprocessing of oleaginous microorganisms

Sonowal

Chikkaputtaiah

Velmurugan

2019

J of Chemical Tech & Biotech

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Oleaginous microorganisms, such as yeast, fungi, microalgae and bacteria, represent a key segment of second generation feed‐stock materials and are considered to synthesize a wide range of industrially important chemical compounds. Oleaginous microorganisms possess a broad varieties of chemical compounds such as carotenoids, pigments, carbohydrates, chlorophyll, and storage‐material lipids. Oleaginous microorganisms have been recognized as promising sources for the synthesis of unsaturated, especially polyunsaturated fatty acids (PUFA). So far, a variety of high‐throughput screening methodologies (HTMs) have been employed for the development of bioprocessing of oleaginous microorganisms for sustainable production of industrially valuable compounds. Of HTMs, flow cytometry (FC) and sorters (FACS) have received substantial interest as better HTMs because of their ability to screen large numbers of cells within seconds, and interrogate and isolate living cells at single‐cell level. Forward and side scattering signals of FC are used to determine the physiological state of the cell while different channels available in the FC facilitate the detection of signals produced from fluorophores. Simultaneous measurement of physiological characteristics along with specific compound accumulation at single‐cell level enables the possibility of separating a particular phenotype with specific properties from a population. Different microbial strain development strategies in combination with FACS produced improved phenotypes with desired properties. This review first summarizes the FACS methodologies suitable for oleaginous microorganisms and the significant progress that has been achieved in oleaginous microorganisms using FACS, and highlights the important, advanced and future prospects of FACS methodologies that are suitable for the development of bioprocessing in oleaginous microorganisms. © 2018 Society of Chemical Industry

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Section: Methods Of Flow Cytometry Suitable For Oleaginous Microorganmentioning

confidence: 99%

Section: Methods Of Flow Cytometry Suitable For Oleaginous Microorganmentioning

confidence: 99%

Section: Methods Of Flow Cytometry Suitable For Oleaginous Microorganmentioning

confidence: 99%

Section: Methods Of Flow Cytometry Suitable For Oleaginous Microorganmentioning

confidence: 99%

Section: Applications Of Flow Cytometry In Oleaginous Microorganismsmentioning

confidence: 99%

See 3 more Smart Citations

Role of flow cytometry for the improvement of bioprocessing of oleaginous microorganisms

Sonowal

Chikkaputtaiah

Velmurugan

2019

J of Chemical Tech & Biotech

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High‐Throughput Raman‐Activated Cell Sorting in the Fingerprint Region

Lindley

Pablo

Peterson

et al. 2022

Adv Materials Technologies

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Cell sorting is the workhorse of biological research and medicine. [1] Cell sorters are commonly used to sort heterogeneous populations of cells based on their intrinsic features and have numerous applications in microbiology, immunology, virology, stem cell biology, and metabolic engineering. [2][3][4][5][6] While several schemes have been developed for sorting cells based on features such as size, [7] morphology, [8][9][10] viability, [11,12] and surface antigens, [13] those able to sort at high throughput for intracellular chemical content are limited to fluorescence-activated and Raman-activated cell sorting (FACS and RACS, respectively). [1,14] In both methods, sorting is accomplished by a stepwise automatic process: 1) measurement of chemical content with single-cell resolution, 2) analysis of the measurement data to identify cells of interest, and 3) mechanical separation of target cells from the original sample. [1] FACS machines commonly identify molecular targets by signals from attached exogenous fluorophores or coexpressed transgenic fluorescent proteins. This indirect detection scheme is often referred to as labeled detection. [1] Although fluorescent labeling is essential for microbiological research, the use of labels has inherrent drawbacks. These include reduced cellular vitality due to the cytotoxicity of labels themselves, nonspecific binding of labels, a lack of labels or transfection methods for many molecular targets and species, incompatibility with human-targeted cell therapies, and increased experimental complexity imposed by labeling protocols. [14] On the other hand, RACS machines directly identify target molecules in a label-free manner via Raman spectroscopy, which measures the inelastic scattering of incident photons by characteristic molecular vibrations. [14] The chemical specificity of this optical measurement contrasts with non-chemically-specific label-free methods such as impedance-based sorting for cell viability and image-based sorting for morphology or deformability. [10][11][12] To date, FACS technology is well developed, commercialized, and broadly adopted, while RACS is nascent and comprised of a small number of lab-based demonstrations. [14][15][16][17][18][19][20][21][22][23][24][25][26] A major hurdle in the broad deployment of RACS for biomedical applications is the weak signal produced by the Raman scattering process. This necessitates comparatively Cell sorting is the workhorse of biological research and medicine. Cell sorters are commonly used to sort heterogeneous cell populations based on their intrinsic features. Raman-activated cell sorting (RACS) has recently received considerable interest by virtue of its ability to discriminate cells by their intracellular chemical content, in a label-free manner. However, the broad deployment of RACS beyond lab-based demonstrations is hindered by a fundamental trade-off between throughput and measurement bandwidth (i.e., cellular information content). Here this trade-off is overcome and broadband RACS in the fingerprint reg...

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Microfluidics for microalgal biotechnology

Özdalgiç

Ustun

Dabbagh

et al. 2021

Biotech & Bioengineering

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Microalgae have expanded their roles as renewable and sustainable feedstocks for biofuel, smart nutrition, biopharmaceutical, cosmeceutical, biosensing, and space technologies. They accumulate valuable biochemical compounds from protein, carbohydrate, and lipid groups, including pigments and carotenoids. Microalgal biomass, which can be adopted for multivalorization under biorefinery settings, allows not only the production of various biofuels but also other value‐added biotechnological products. However, state‐of‐the‐art technologies are required to optimize yield, quality, and the economical aspects of both upstream and downstream processes. As such, the need to use microfluidic‐based devices for both fundamental research and industrial applications of microalgae, arises due to their microscale sizes and dilute cultures. Microfluidics‐based devices are superior to their competitors through their ability to perform multiple functions such as sorting and analyzing small amounts of samples (nanoliter to picoliter) with higher sensitivities. Here, we review emerging applications of microfluidic technologies on microalgal processes in cell sorting, cultivation, harvesting, and applications in biofuels, biosensing, drug delivery, and nutrition.

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Fluorescence activated cell-sorting principles and applications in microalgal biotechnology

Cited by 68 publications

References 64 publications

Role of flow cytometry for the improvement of bioprocessing of oleaginous microorganisms

Role of flow cytometry for the improvement of bioprocessing of oleaginous microorganisms

High‐Throughput Raman‐Activated Cell Sorting in the Fingerprint Region

Microfluidics for microalgal biotechnology

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