Authors

Lee, S. S., Avalos Vizcarra, I., Huberts, D. H. E. W., Lee, L. P., & Heinemann, M.

Affiliations

A Institute of Molecular Systems Biology, ETH Zurich, 8093 Zurich, Switzerland; b Department of Biosystems Science and Engineering, ETH Zurich, 4058 Basel, Switzerland; and c Molecular Systems Biology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, 9747 AG Groningen, The Netherlands.

Application Area

General Biology

General Fluorescence Microscopy

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.

Extract

… “automatically and stably maintained the focus during the whole experiment. LED (pE2; CoolLed) illumination was used as the fluorescent light source.”…

Product Associated Features

pE-2: A repeatable, controllable modular system with 20 different LED peaks. Instant on/off and intensity (0-100%) control.

Product Type

-

Journal

PNAS

Year of Publication

2012

Country of Publication

Switzerland