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Title: Compartmentalized microchannel array for high-throughput analysis of single cell polarized growth and dynamics

Abstract

Here, interrogating polarized growth is technologically challenging due to extensive cellular branching and uncontrollable environmental conditions in conventional assays. Here we present a robust and high-performance microfluidic system that enables observations of polarized growth with enhanced temporal and spatial control over prolonged periods. The system has built-in tunability and versatility to accommodate a variety of science applications requiring precisely controlled environments. Using the model filamentous fungus, Neurospora crassa, this microfluidic system enabled direct visualization and analysis of cellular heterogeneity in a clonal fungal cell population, nuclear distribution and dynamics at the subhyphal level, and quantitative dynamics of gene expression with single hyphal compartment resolution in response to carbon source starvation and exchange experiments. Although the microfluidic device is demonstrated on filamentous fungi, our technology is immediately extensible to a wide array of other biosystems that exhibit similar polarized cell growth with applications ranging from bioenergy production to human health.

Authors:
 [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1]
  1. Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER)
OSTI Identifier:
1226407
Report Number(s):
PNNL-SA-109268
Journal ID: ISSN 2045-2322; 48672; KP1704020
Grant/Contract Number:  
AC05-76RL01830; 66382
Resource Type:
Accepted Manuscript
Journal Name:
Scientific Reports
Additional Journal Information:
Journal Volume: 5; Journal Issue: 16111; Journal ID: ISSN 2045-2322
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; microchannel; high-throughput; cell; polarized growth; microfluidic; Neurospora crassa; Environmental Molecular Sciences Laboratory; fungal biology; lab-on-a-chip

Citation Formats

Geng, Tao, Bredeweg, Erin L., Szymanski, Craig J., Liu, Bingwen, Baker, Scott E., Orr, Galya, Evans, James E., and Kelly, Ryan T. Compartmentalized microchannel array for high-throughput analysis of single cell polarized growth and dynamics. United States: N. p., 2015. Web. doi:10.1038/srep16111.
Geng, Tao, Bredeweg, Erin L., Szymanski, Craig J., Liu, Bingwen, Baker, Scott E., Orr, Galya, Evans, James E., & Kelly, Ryan T. Compartmentalized microchannel array for high-throughput analysis of single cell polarized growth and dynamics. United States. doi:10.1038/srep16111.
Geng, Tao, Bredeweg, Erin L., Szymanski, Craig J., Liu, Bingwen, Baker, Scott E., Orr, Galya, Evans, James E., and Kelly, Ryan T. Wed . "Compartmentalized microchannel array for high-throughput analysis of single cell polarized growth and dynamics". United States. doi:10.1038/srep16111. https://www.osti.gov/servlets/purl/1226407.
@article{osti_1226407,
title = {Compartmentalized microchannel array for high-throughput analysis of single cell polarized growth and dynamics},
author = {Geng, Tao and Bredeweg, Erin L. and Szymanski, Craig J. and Liu, Bingwen and Baker, Scott E. and Orr, Galya and Evans, James E. and Kelly, Ryan T.},
abstractNote = {Here, interrogating polarized growth is technologically challenging due to extensive cellular branching and uncontrollable environmental conditions in conventional assays. Here we present a robust and high-performance microfluidic system that enables observations of polarized growth with enhanced temporal and spatial control over prolonged periods. The system has built-in tunability and versatility to accommodate a variety of science applications requiring precisely controlled environments. Using the model filamentous fungus, Neurospora crassa, this microfluidic system enabled direct visualization and analysis of cellular heterogeneity in a clonal fungal cell population, nuclear distribution and dynamics at the subhyphal level, and quantitative dynamics of gene expression with single hyphal compartment resolution in response to carbon source starvation and exchange experiments. Although the microfluidic device is demonstrated on filamentous fungi, our technology is immediately extensible to a wide array of other biosystems that exhibit similar polarized cell growth with applications ranging from bioenergy production to human health.},
doi = {10.1038/srep16111},
journal = {Scientific Reports},
number = 16111,
volume = 5,
place = {United States},
year = {2015},
month = {11}
}

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