scholarly journals Single Cell Analysis of Bistable Expression of Pathogenicity Island 1 and the Flagellar Regulon in Salmonella enterica

2021 ◽  
Vol 9 (2) ◽  
pp. 210
Author(s):  
María Antonia Sánchez-Romero ◽  
Josep Casadesús
Keyword(s):  
Cell Sorting ◽  
Salmonella Enterica ◽  
Single Cell Analysis ◽  
Pathogenicity Island ◽  
Time Lapse ◽  
Cell Analysis ◽  
Microscopy Observation ◽  
Time Lapse Microscopy ◽  
Flagellar Regulon ◽  
Invasion Assays

Bistable expression of the Salmonella enterica pathogenicity island 1 (SPI-1) and the flagellar network (Flag) has been described previously. In this study, simultaneous monitoring of OFF and ON states in SPI-1 and in the flagellar regulon reveals independent switching, with concomitant formation of four subpopulations: SPI-1OFF FlagOFF, SPI-1OFF FlagON, SPI-1ON FlagOFF, and SPI-1ON FlagON. Invasion assays upon cell sorting show that none of the four subpopulations is highly invasive, thus raising the possibility that FlagOFF cells might contribute to optimal invasion as previously proposed for SPI-1OFF cells. Time lapse microscopy observation indicates that expression of the flagellar regulon contributes to the growth impairment previously described in SPI-1ON cells. As a consequence, growth resumption in SPI-1ON FlagON cells requires switching to both SPI-1OFF and FlagOFF states.

New Micro Devices for Single Cell Analysis, Cell Sorting and Cloning-on-a-Chip: The CytoconTM Instrument

2000 ◽  
pp. 443-446 ◽  
Author(s):  
G. Gradl ◽  
T. Müller ◽  
A. Pfennig ◽  
S. Shirley ◽  
T. Schnelle ◽  
...  
Keyword(s):  
Single Cell ◽  
Cell Sorting ◽  
Single Cell Analysis ◽  
Cell Analysis

scholarly journals Reporters for Single-Cell Analysis of Colicin Ib Expression in Salmonella enterica Serovar Typhimurium

PLoS ONE ◽  
2015 ◽  
Vol 10 (12) ◽  
pp. e0144647 ◽  
Author(s):  
Stefanie Spriewald ◽  
Jana Glaser ◽  
Markus Beutler ◽  
Martin B. Koeppel ◽  
Bärbel Stecher
Keyword(s):  
Single Cell ◽  
Salmonella Enterica ◽  
Salmonella Enterica Serovar Typhimurium ◽  
Single Cell Analysis ◽  
Cell Analysis ◽  
Serovar Typhimurium

Time lapse microscopy observation of cellular structural changes and image analysis of drug treated cancer cells to characterize the cellular heterogeneity

2014 ◽  
Vol 30 (6) ◽  
pp. 724-734 ◽  
Author(s):  
Periasamy S. Vaiyapuri ◽  
Alshatwi A. Ali ◽  
Akbarsha A. Mohammad ◽  
Jeyalakshmi Kandhavelu ◽  
Meenakshisundaram Kandhavelu
Keyword(s):  
Image Analysis ◽  
Cancer Cells ◽  
Structural Changes ◽  
Time Lapse ◽  
Cellular Heterogeneity ◽  
Microscopy Observation ◽  
Time Lapse Microscopy

scholarly journals Isolation and Time Lapse Microscopy of Highly Pure Hepatic Stellate Cells

2015 ◽  
Vol 2015 ◽  
pp. 1-13 ◽  
Author(s):  
Matthias Bartneck ◽  
Klaudia Theresa Warzecha ◽  
Carmen Gabriele Tag ◽  
Sibille Sauer-Lehnen ◽  
Felix Heymann ◽  
...  
Keyword(s):  
Liver Fibrosis ◽  
Hepatic Stellate Cells ◽  
Cell Sorting ◽  
Stellate Cells ◽  
Time Lapse ◽  
Functional Responses ◽  
Effector Cells ◽  
Time Lapse Microscopy ◽  
Density Centrifugation

Hepatic stellate cells (HSC) are the main effector cells for liver fibrosis. We aimed at optimizing HSC isolation by an additional step of fluorescence-activated cell sorting (FACS) via a UV laser. HSC were isolated from livers of healthy mice and animals subjected to experimental fibrosis. HSC isolation by iohexol- (Nycodenz) based density centrifugation was compared to a method with subsequent FACS-based sorting. We assessed cellular purity, viability, morphology, and functional properties like proliferation, migration, activation marker, and collagen expression. FACS-augmented isolation resulted in a significantly increased purity of stellate cells (>99%) compared to iohexol-based density centrifugation (60–95%), primarily by excluding doublets of HSC and Kupffer cells (KC). Importantly, this method is also applicable to young animals and mice with liver fibrosis. Viability, migratory properties, and HSC transdifferentiationin vitrowere preserved upon FACS-based isolation, as assessed using time lapse microscopy. During maturation of HSC in culture, we did not observe HSC cell division using time lapse microscopy. Strikingly, FACS-isolated, differentiated HSC showed very limited molecular and functional responses to LPS stimulation. In conclusion, isolating HSC from mouse liver by additional FACS significantly increases cell purity by removing contaminations from other cell populations especially KC, without affecting HSC viability, migration, or differentiation.

scholarly journals Cyto-Mine: An Integrated, Picodroplet System for High-Throughput Single-Cell Analysis, Sorting, Dispensing, and Monoclonality Assurance

2020 ◽  
Vol 25 (2) ◽  
pp. 177-189 ◽  
Author(s):  
Dimitris Josephides ◽  
Serena Davoli ◽  
William Whitley ◽  
Raphael Ruis ◽  
Robert Salter ◽  
...  
Keyword(s):  
Single Cell ◽  
Cell Line ◽  
High Throughput ◽  
Cell Sorting ◽  
High Throughput Screening ◽  
Single Cell Analysis ◽  
Microtiter Plate ◽  
Cell Analysis ◽  
Fully Integrated ◽  
Clonal Cell Lines

The primary goal of bioprocess cell line development is to obtain high product yields from robustly growing and well-defined clonal cell lines in timelines measured in weeks rather than months. Likewise, high-throughput screening of B cells and hybridomas is required for most cell line engineering workflows. A substantial bottleneck in these processes is detecting and isolating rare clonal cells with the required characteristics. Traditionally, this was achieved by the resource-intensive method of limiting dilution cloning, and more recently aided by semiautomated technologies such as cell sorting (e.g., fluorescence-activated cell sorting) and colony picking. In this paper we report on our novel Cyto-Mine Single Cell Analysis and Monoclonality Assurance System, which overcomes the limitations of current technologies by screening hundreds of thousands of individual cells for secreted target proteins, and then isolating and dispensing the highest producers into microtiter plate wells (MTP). The Cyto-Mine system performs this workflow using a fully integrated, microfluidic Cyto-Cartridge. Critically, all reagents and Cyto-Cartridges used are animal component-free (ACF) and sterile, thus allowing fast, robust, and safe isolation of desired cells.

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