scholarly journals Single-cell microscopy of suspension cultures using a microfluidics-assisted cell screening platform

2017 ◽  
Vol 13 (1) ◽  
pp. 170-194 ◽  
Author(s):  
Burak Okumus ◽  
Charles J Baker ◽  
Juan Carlos Arias-Castro ◽  
Ghee Chuan Lai ◽  
Emanuele Leoncini ◽  
...  
Keyword(s):  
Single Cell ◽  
Suspension Cultures ◽  
Screening Platform ◽  
Cell Microscopy

scholarly journals FleN contributes to heterogeneous swimming at high temperatures inPseudomonas syringae

2018 ◽  
Author(s):  
Kevin L. Hockett ◽  
Steven E. Lindow
Keyword(s):  
Flow Cytometry ◽  
Single Cell ◽  
Pseudomonas Syringae ◽  
Environmental Conditions ◽  
Promoter Region ◽  
Plant Pathogens ◽  
Incubation Temperature ◽  
Nutrient Depletion ◽  
Synonymous Mutations ◽  
Cell Microscopy

SUMMARYMotility is generally conserved among many animal and plant pathogens. Environmental conditions, however, significantly impact expression of the motile phenotype. In this study, we describe a novel heterogeneous motility phenotype inPseudomonas syringae, where under normally suppressive incubation conditions (30°C) punctate colonies arise that are spatially isolated from the point of inoculation, giving rise to a motility pattern we term constellation swimming (CS). We demonstrate that this phenotype is reproducible, reversible, and dependent on a functioning flagellum. Mirroring the heterogeneous motility phenotype, we demonstrate the existence of a sub-population of cells under non-permissive conditions that express flagellin (fliC) at levels similar to cells incubated under permissive conditions using both quantitative single cell microscopy and flow cytometry. To understand the genetics underlying the CS phenotype, we selected for naturally arising mutants that exhibited a normal swimming phenotype at the warmer incubation temperature. Sequencing these mutants recovered several independent non-synonymous mutations within FleN (also known as FlhG) as well as mutations within the promoter region of FleQ, the master flagellum regulator inPseudomonas. We further show that nutrient depletion is the likely underlying cause of CS, as reduced nutrients will stimulate bothfliCexpression and a normal swimming phenotype at 30 °C.

scholarly journals A novel single-cell screening platform reveals proteome plasticity during yeast stress responses

2013 ◽  
Vol 200 (6) ◽  
pp. 839-850 ◽  
Author(s):  
Michal Breker ◽  
Melissa Gymrek ◽  
Maya Schuldiner
Keyword(s):  
Single Cell ◽  
Stress Responses ◽  
Nitrogen Starvation ◽  
Bet Hedging ◽  
Transcriptional Responses ◽  
Cellular Physiology ◽  
Protein Levels ◽  
Hedging Strategies ◽  
Screening Platform ◽  
External Stimuli

Uncovering the mechanisms underlying robust responses of cells to stress is crucial for our understanding of cellular physiology. Indeed, vast amounts of data have been collected on transcriptional responses in Saccharomyces cerevisiae. However, only a handful of pioneering studies describe the dynamics of proteins in response to external stimuli, despite the fact that regulation of protein levels and localization is an essential part of such responses. Here we characterized unprecedented proteome plasticity by systematically tracking the localization and abundance of 5,330 yeast proteins at single-cell resolution under three different stress conditions (DTT, H2O2, and nitrogen starvation) using the GFP-tagged yeast library. We uncovered a unique “fingerprint” of changes for each stress and elucidated a new response arsenal for adapting to radical environments. These include bet-hedging strategies, organelle rearrangement, and redistribution of protein localizations. All data are available for download through our online database, LOQATE (localization and quantitation atlas of yeast proteome).

Investigation of selective release of periplasmic proteins through pore size analysis and single-cell microscopy in Escherichia coli

2021 ◽  
pp. 108009
Author(s):  
Subbarayalu Ramalakshmi ◽  
Ramakrishnan Nagasundara Ramanan ◽  
Shanmugavel Madhavan ◽  
Chien Wei Ooi ◽  
Catherine Ching Han Chang ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Pore Size ◽  
Single Cell ◽  
Size Analysis ◽  
Pore Size Analysis ◽  
Periplasmic Proteins ◽  
Cell Microscopy

scholarly journals Subcellular functions of proteins under fluorescence single-cell microscopy

2016 ◽  
Vol 1864 (1) ◽  
pp. 77-84 ◽  
Author(s):  
Casey L. Kohnhorst ◽  
Danielle L. Schmitt ◽  
Anand Sundaram ◽  
Songon An
Keyword(s):  
Single Cell ◽  
Cell Microscopy
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