Microfluidics device for single cell gene expression analysis in Saccharomyces cerevisiae

@article{Ryley2006MicrofluidicsDF,
  title={Microfluidics device for single cell gene expression analysis in Saccharomyces cerevisiae},
  author={James Ryley and Olivia Pereira-smith},
  journal={Yeast},
  year={2006},
  volume={23}
}
We have measured single‐cell gene expression over time using a microfluidics‐based flow cell which physically traps individual yeast using µm‐sized structures (yeast jails). Our goal was to determine variability of gene expression within a cell over time, as well as variability between individual cells. In our flow cell system, yeast jails are fabricated out of PDMS and gene expression is visualized using fluorescently‐tagged proteins of interest. Previously, single‐cell yeast work has been… 

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References

SHOWING 1-10 OF 35 REFERENCES

Transport, retention and fluorescent measurement of single biological cells studied in microfluidic chips.

It is demonstrated that Jurkat cells remain viable within microfluidic channels for cellular studies and that the microfluidity chip can facilitate monitoring of cellular changes of biological cells at the single cell level and in the same cell.

Single-cell enzyme concentrations, kinetics, and inhibition analysis using high-density hydrodynamic cell isolation arrays.

This technology was applied to determine novel single-cell enzyme kinetics for three different cell types (HeLa, 293T, Jurkat) and predicted this varied response was due to variation in the concentration of carboxylesterase between cell types.

Microfabricated platform for studying stem cell fates

A simple, versatile, and efficient micropatterned arraying system conducive to the culture and dynamic monitoring of stem cell proliferation and further interrogate the response of distinct stem cell subpopulations to microenvironmental cues that govern their behavior.

Microfluidic device for single-cell analysis.

A novel microfluidic device constructed from poly(dimethylsiloxane) using multilayer soft lithography technology for the analysis of single cells is developed, achieving significant improvements in reagent consumption, analysis time, and temporal resolution over macroscale alternatives.

Heterogeneity of stress gene expression and stress resistance among individual cells of Saccharomyces cerevisiae

Flow cytometry and fluorescence techniques indicated that, although the mean gene expression by a population is influenced by the genetic background, the heterogeneity among individual cells in clonal populations is largely physiologically based.

Microfluidic selection and retention of a single cardiac myocyte, on-chip dye loading, cell contraction by chemical stimulation, and quantitative fluorescent analysis of intracellular calcium.

A microfluidic method to study the contraction of a single cardiac myocyte (heart muscle cell) has been developed. This method integrates various single-cell operations as well as on-chip dye

Large-scale single-cell trapping and imaging using microwell arrays.

An optimization study for trapping single cells with high efficiency in large arrays of microwells and characterized microwell occupancy by cells for a range of microwell dimensions and seeding parameters and optimized it for fibroblasts and rat basophilic leukemia cells.

Regulation of noise in the expression of a single gene

The results provide the first direct experimental evidence of the biochemical origin of phenotypesic noise, demonstrating that the level of phenotypic variation in an isogenic population can be regulated by genetic parameters.

Microarray analysis of replicative senescence

Real‐time imaging of transcriptional activation in live cells reveals rapid up‐regulation of the cyclin‐dependent kinase inhibitor gene CDKN1A in replicative cellular senescence

A new in vitro human model system that allows entry into senescence to be monitored in real‐time in individual viable cells is reported here and shows its proper control by physiological signalling circuits.