Flow-cytometry and cell sorting: An efficient approach to investigate productivity and cell physiology in mammalian cell factories

Methods ◽  
2012 ◽  
Vol 56 (3) ◽  
pp. 366-374 ◽  
Author(s):  
Niraj Kumar ◽  
Nicole Borth
Keyword(s):  
Flow Cytometry ◽  
Mammalian Cell ◽  
Cell Sorting ◽  
Cell Physiology ◽  
Efficient Approach ◽  
Cell Factories

scholarly journals Multiparametric Flow Cytometry and Cell Sorting for the Assessment of Viable, Injured, and Dead Bifidobacterium Cells during Bile Salt Stress

2002 ◽  
Vol 68 (11) ◽  
pp. 5209-5216 ◽  
Author(s):  
Kaouther Ben Amor ◽  
Pieter Breeuwer ◽  
Patrick Verbaarschot ◽  
Frank M. Rombouts ◽  
Antoon D. L. Akkermans ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Salt Stress ◽  
Bile Salt ◽  
Cell Sorting ◽  
Physiological Activity ◽  
Membrane Integrity ◽  
Multiparameter Flow Cytometry ◽  
Bifidobacterium Adolescentis ◽  
Multiparametric Flow Cytometry ◽  
Plate Counts

ABSTRACT Using a flow cytometry-based approach, we assessed the viability of Bifidobacterium lactis DSM 10140 and Bifidobacterium adolescentis DSM 20083 during exposure to bile salt stress. Carboxyfluorescein diacetate (cFDA), propidium iodide (PI), and oxonol [DiBAC4(3)] were used to monitor esterase activity, membrane integrity, and membrane potential, respectively, as indicators of bacterial viability. Single staining with these probes rapidly and noticeably reflected the behavior of the two strains during stress exposure. However, the flow cytometry results tended to overestimate the viability of the two strains compared to plate counts, which appeared to be related to the nonculturability of a fraction of the population as a result of sublethal injury caused by bile salts. When the cells were simultaneously stained with cFDA and PI, flow cytometry and cell sorting revealed a striking physiological heterogeneity within the stressed bifidobacterium population. Three subpopulations could be identified based on their differential uptake of the probes: cF-stained, cF and PI double-stained, and PI-stained subpopulations, representing viable, injured, and dead cells, respectively. Following sorting and recovery, a significant fraction of the double-stained subpopulation (40%) could resume growth on agar plates. Our results show that in situ assessment of the physiological activity of stressed bifidobacteria using multiparameter flow cytometry and cell sorting may provide a powerful and sensitive tool for assessment of the viability and stability of probiotics.

Flow cytometry and cell sorting for yeast viability assessment and cell selection

Yeast ◽  
1998 ◽  
Vol 14 (2) ◽  
pp. 147-160 ◽  
Author(s):  
Daniel Deere ◽  
Jian Shen ◽  
Graham Vesey ◽  
Philip Bell ◽  
Peter Bissinger ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Cell Sorting ◽  
Cell Selection ◽  
Viability Assessment ◽  
Yeast Viability

scholarly journals High-throughput screening of high lactic acid-producing Bacillus coagulans by droplet microfluidic based flow cytometry with fluorescence activated cell sorting

RSC Advances ◽  
2019 ◽  
Vol 9 (8) ◽  
pp. 4507-4513 ◽  
Author(s):  
Xu-Dong Zhu ◽  
Xiang Shi ◽  
Shu-Wen Wang ◽  
Ju Chu ◽  
Wei-Hong Zhu ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Lactic Acid ◽  
High Throughput ◽  
Cell Sorting ◽  
High Throughput Screening ◽  
Bacillus Coagulans ◽  
Screening System ◽  
Fluorescence Activated Cell Sorting ◽  
Microfluidic Sorting

A high-throughput screening system based on droplet microfluidic sorting was developed and employed for screening of high lactic acid-producing Bacillus coagulans.

scholarly journals Improvement of phytoplankton culture isolation using single cell sorting by flow cytometry

2017 ◽  
Vol 53 (2) ◽  
pp. 271-282 ◽  
Author(s):  
Dominique Marie ◽  
Florence Le Gall ◽  
Roseline Edern ◽  
Priscillia Gourvil ◽  
Daniel Vaulot
Keyword(s):  
Flow Cytometry ◽  
Single Cell ◽  
Cell Sorting
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