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Proc Natl Acad Sci U S A. 2012 Mar 27;109(13):4916-20. doi: 10.1073/pnas.1113505109. Epub 2012 Mar 14.

Whole lifespan microscopic observation of budding yeast aging through a microfluidic dissection platform.

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1
Institute of Molecular Systems Biology, ETH Zurich, 8093 Zurich, Switzerland.

Abstract

Important insights into aging have been generated with the genetically tractable and short-lived budding yeast. However, it is still impossible today to continuously track cells by high-resolution microscopic imaging (e.g., fluorescent imaging) throughout their entire lifespan. Instead, the field still needs to rely on a 50-y-old laborious and time-consuming method to assess the lifespan of yeast cells and to isolate differentially aged cells for microscopic snapshots via manual dissection of daughter cells from the larger mother cell. Here, we are unique in achieving continuous and high-resolution microscopic imaging of the entire replicative lifespan of single yeast cells. Our microfluidic dissection platform features an optically prealigned single focal plane and an integrated array of soft elastomer-based micropads, used together to allow for trapping of mother cells, removal of daughter cells, monitoring gradual changes in aging, and unprecedented microscopic imaging of the whole aging process. Using the platform, we found remarkable age-associated changes in phenotypes (e.g., that cells can show strikingly differential cell and vacuole morphologies at the moment of their deaths), indicating substantial heterogeneity in cell aging and death. We envision the microfluidic dissection platform to become a major tool in aging research.

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PMID:
22421136
PMCID:
PMC3324001
DOI:
10.1073/pnas.1113505109
[Indexed for MEDLINE]
Free PMC Article
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