TY - JOUR
T1 - High-throughput live imaging of microcolonies to measure heterogeneity in growth and gene expression
AU - Sartori, Federica M.O.
AU - Buzby, Cassandra
AU - Plavskin, Yevgeniy
AU - Siegal, Mark L.
N1 - Funding Information:
We thank Naomi Ziv, Sasha Levy and Shuang Li for their contributions to developing this protocol, David Gresham for shared equipment, and Marissa Knoll for help with video production. This work was supported by National Institutes of Health grant R35GM118170.
Publisher Copyright:
© 2021 JoVE Journal of Visualized Experiments.
PY - 2021/4
Y1 - 2021/4
N2 - Precise measurements of between-and within-strain heterogeneity in microbial growth rates are essential for understanding genetic and environmental inputs into stress tolerance, pathogenicity, and other key components of fitness. This manuscript describes a microscope-based assay that tracks approximately 105 Saccharomyces cerevisiae microcolonies per experiment. After automated time-lapse imaging of yeast immobilized in a multiwell plate, microcolony growth rates are easily analyzed with custom image-analysis software. For each microcolony, expression and localization of fluorescent proteins and survival of acute stress can also be monitored. This assay allows precise estimation of strains' average growth rates, as well as comprehensive measurement of heterogeneity in growth, gene expression, and stress tolerance within clonal populations.
AB - Precise measurements of between-and within-strain heterogeneity in microbial growth rates are essential for understanding genetic and environmental inputs into stress tolerance, pathogenicity, and other key components of fitness. This manuscript describes a microscope-based assay that tracks approximately 105 Saccharomyces cerevisiae microcolonies per experiment. After automated time-lapse imaging of yeast immobilized in a multiwell plate, microcolony growth rates are easily analyzed with custom image-analysis software. For each microcolony, expression and localization of fluorescent proteins and survival of acute stress can also be monitored. This assay allows precise estimation of strains' average growth rates, as well as comprehensive measurement of heterogeneity in growth, gene expression, and stress tolerance within clonal populations.
UR - http://www.scopus.com/inward/record.url?scp=85105887242&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85105887242&partnerID=8YFLogxK
U2 - 10.3791/62038
DO - 10.3791/62038
M3 - Article
C2 - 33938878
AN - SCOPUS:85105887242
VL - 2021
JO - Journal of Visualized Experiments
JF - Journal of Visualized Experiments
SN - 1940-087X
IS - 170
M1 - e62038
ER -