scholarly journals FACS Single Cell Sorting v3 (protocols.io.besbjean)

protocols.io ◽  
2020 ◽  
Author(s):  
Allen Institute
Keyword(s):  
Single Cell ◽  
Cell Sorting

scholarly journals Gezielte Zellsortierung in der Einzelzellgenomik

BIOspektrum ◽  
2021 ◽  
Vol 27 (3) ◽  
pp. 274-276
Author(s):  
Morgan S. Sobol ◽  
Anne-Kristin Kaster
Keyword(s):  
Dark Matter ◽  
Single Cell ◽  
Cell Sorting ◽  
Genetic Information ◽  
Fluorescent Labeling ◽  
Single Cell Genomics ◽  
Rare Biosphere ◽  
Prokaryotic Genomes ◽  
Selection Of

AbstractSingle cell genomics (SCG) can provide reliable context for assembled genome fragments on the level of individual prokaryotic genomes and has rapidly emerged as an essential complement to cultivation-based and metagenomics research approaches. Targeted cell sorting approaches, which enable the selection of specific taxa by fluorescent labeling, compatible with subsequent single cell genomics offers an opportunity to access genetic information from rare biosphere members which would have otherwise stayed hidden as microbial dark matter.

Hapten-Specific Single-Cell Selection of Hybridoma Clones by Fluorescence-Activated Cell Sorting for the Generation of Monoclonal Antibodies

2017 ◽  
Vol 89 (7) ◽  
pp. 4007-4012 ◽  
Author(s):  
Martin Dippong ◽  
Peter Carl ◽  
Christine Lenz ◽  
Jörg A. Schenk ◽  
Katrin Hoffmann ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Single Cell ◽  
Cell Sorting ◽  
Cell Selection ◽  
Fluorescence Activated Cell Sorting ◽  
Selection Of

There are no A1 and A2 astrocytes in brain

2021 ◽  
Author(s):  
Shuang-qi Gao
Keyword(s):  
Endothelial Cells ◽  
Single Cell ◽  
Rna Sequencing ◽  
Cell Sorting ◽  
Reactive Astrocytes ◽  
Marker Genes ◽  
Set Up ◽  
The Brain

Abstract Objectives The subsets of astrocytes in the brain have not been fully elucidated. Using bulk RNA sequencing, reactive astrocytes were divided into A1 versus A2. However, using single-cell RNAseq (ScRNAseq), astrocytes were divided into over two subsets. Our aim was to set up the correspondence between the fluorescent-activated cell sorting (FACS)-bulk RNAseq and ScRNAseq data. Results We found that most of reactive astrocytes (RAs) marker genes were expressed in endothelial cells but not in astrocytes, suggesting those marker genes are not suitable for astrocytic activation. The absence of A1 and A2 astrocytes in the brain.

scholarly journals Click-chemistry enabled directed evolution of glycosynthases for bespoke glycans synthesis

2020 ◽  
Author(s):  
Ayushi Agrawal ◽  
Chandra Kanth Bandi ◽  
Tucker Burgin ◽  
Youngwoo Woo ◽  
Heather B. Mayes ◽  
...  
Keyword(s):  
Single Cell ◽  
Click Chemistry ◽  
Directed Evolution ◽  
Cell Sorting ◽  
Screening Methods ◽  
Carbohydrate Active Enzymes ◽  
E Coli ◽  
Highly Sensitive ◽  
Increased Activity

AbstractEngineering of carbohydrate-active enzymes like glycosynthases for chemoenzymatic synthesis of bespoke oligosaccharides has been limited by the lack of suitable directed evolution based protein engineering methods. Currently there are no ultrahigh-throughput screening methods available for rapid and highly sensitive single cell-based screening of evolved glycosynthase enzymes employing azido sugars as substrates. Here, we report a fluorescence-based approach employing click-chemistry for the selective detection of glycosyl azides (versus free inorganic azides) that facilitated ultrahigh-throughput in-vivo single cell-based assay of glycosynthase activity. This discovery has led to the development of a directed evolution methodology for screening and sorting glycosynthase mutants for synthesis of desired fucosylated oligosaccharides. Our screening technique facilitated rapid fluorescence activated cell sorting of a large library of glycosynthase variants (>106 mutants) expressed in E. coli to identify several novel mutants with increased activity for β-fucosyl-azide activated donor sugars towards desired acceptor sugars, demonstrating the broader applicability of this methodology.

scholarly journals Large particle fluorescence-activated cell sorting enables high quality single cell RNA-sequencing and functional analysis of adult cardiomyocytes

2019 ◽  
Author(s):  
Suraj Kannan ◽  
Matthew Miyamoto ◽  
Brian Lin ◽  
Renjun Zhu ◽  
Sean Murphy ◽  
...  
Keyword(s):  
Single Cell ◽  
Rna Sequencing ◽  
Cell Sorting ◽  
Large Particle ◽  
Fluorescence Activated Cell Sorting ◽  
High Quality ◽  
Adult Cardiomyocytes ◽  
Single Cell Rna Sequencing ◽  
Mitochondrial Transcripts ◽  
Manual Methods

ABSTRACTRationaleSingle cell RNA sequencing (scRNA-seq) has emerged as a powerful tool to profile the transcriptome at single cell resolution, enabling comprehensive analysis of cellular trajectories and heterogeneity during development and disease. However, the use of scRNA-seq remains limited in cardiac pathology owing to technical difficulties associated with the isolation of single adult cardiomyocytes (CMs).ObjectiveWe investigated the capability of large-particle fluorescence-activated cell sorting (LP-FACS) for isolation of viable single adult CMs.Methods and ResultsWe found that LP-FACS readily outperforms conventional FACS for isolation of struturally competent CMs, including large CMs. Additionally, LP-FACS enables isolation of fluorescent CMs from mosaic models. Importantly, the sorted CMs allow generation of high-quality scRNA-seq libraries. Unlike CMs isolated via previously utilized fluidic or manual methods, LP-FAC-isolated CMs generate libraries exhibiting normal levels of mitochondrial transcripts. Moreover, LP-FACS isolated CMs remain functionally competent and can be studied for contractile properties.ConclusionsOur study enables high quality dissection of adult CM biology at single-cell resolution.

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