Monitoring dynamics of single-cell gene expression over multiple cell cycles.
Recent progress in reconstructing gene regulatory networks has established a framework for a quantitative description of the dynamics of many important cellular processes. Such a description will require novel experimental techniques that enable the generation of time-series data for the governing regulatory proteins in a large number of individual living cells. Here, we utilize microfabrication to construct a Tesla microchemostat that permits single-cell fluorescence imaging of gene expression over many cellular generations. The device is used to capture and constrain asymmetrically dividing or motile cells within a trapping region and to deliver nutrients and regulate the cellular population within this region. We illustrate the operation of the microchemostat with Saccharomyces cerevisiae and explore the evolution of single-cell gene expression and cycle time as a function of generation. Our findings highlight the importance of novel assays for quantifying the dynamics of gene expression and cellular growth, and establish a methodology for exploring the effects of gene expression on long-term processes such as cellular aging.
Cookson
Scott
S
Department of Bioengineering, University of California at San Diego, La Jolla, CA, USA.
Ostroff
Natalie
N
Pang
Wyming Lee
WL
Volfson
Dmitri
D
Hasty
Jeff
J
D001426:Bacterial Proteins / Q000032:analysis; D002453:Cell Cycle; D002465:Cell Movement; D004867:Equipment Design; D015870:Gene Expression; D020869:Gene Expression Profiling / Q000295:instrumentation / Q000379:methods*; D008164:Luminescent Proteins / Q000032:analysis; D008832:Microchemistry; D008856:Microscopy, Fluorescence / Q000295:instrumentation* / Q000379:methods; D008863:Microspheres; D009172:Mycology / Q000295:instrumentation / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012212:Rheology; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis* / Q000235:genetics; D012984:Software; D000097573:Red Fluorescent Protein
D001426:Bacterial Proteins; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D000097573:Red Fluorescent Protein; C054562:yellow fluorescent protein, Bacteria