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  4. Long-Term Single Cell Analysis of S. pombe on a Microfluidic Microchemostat Array
 
research article

Long-Term Single Cell Analysis of S. pombe on a Microfluidic Microchemostat Array

Nobs, Jean-Bernard  
•
Maerkl, Sebastian J.  
2014
Plos One

Although Schyzosaccharomyces pombe is one of the principal model organisms for studying the cell cycle, surprisingly few methods have characterized S. pombe growth on the single cell level, and no methods exist capable of analyzing thousands of cells and tens of thousands of cell division events. We developed an automated microfluidic platform permitting S. pombe to be grown on-chip for several days under defined and changeable conditions. We developed an image processing pipeline to extract and quantitate several physiological parameters including cell length, time to division, and elongation rate without requiring synchronization of the culture. Over a period of 50 hours our platform analyzed over 100000 cell division events and reconstructed single cell lineages up to 10 generations in length. We characterized cell lengths and division times in a temperature shift experiment in which cells were initially grown at 30 degrees C and transitioned to 25 degrees C. Although cell length was identical at both temperatures at steady-state, we observed transient changes in cell length if the temperature shift took place during a critical phase of the cell cycle. We further show that cells born with normal length do divide over a wide range of cell lengths and that cell length appears to be controlled in the second generation, were large newly born cells have a tendency to divide more rapidly and thus at a normalized cell size. The platform is thus applicable to measure fine-details in cell cycle dynamics, should be a useful tool to decipher the molecular mechanism underlying size homeostasis, and will be generally applicable to study processes on the single cell level that require large numbers of precision measurements and single cell lineages.

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Type
research article
DOI
10.1371/journal.pone.0093466
Web of Science ID

WOS:000334159800035

Author(s)
Nobs, Jean-Bernard  
Maerkl, Sebastian J.  
Date Issued

2014

Publisher

Public Library of Science

Published in
Plos One
Volume

9

Issue

4

Article Number

e93466

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
LBNC  
Available on Infoscience
May 19, 2014
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/103446
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