Continuous live cell imaging of cellulose attachment by microbes under anaerobic and thermophilic conditions using confocal microscopy

Zhi Wu Wang, Seung Hwan Lee, James G. Elkins, Yongchao Li, Scott Hamilton-Brehm, Jennifer L. Morrell-Falvey

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

Live cell imaging methods provide important insights into the dynamics of cellular processes that cannot be derived easily from population-averaged datasets. In the bioenergy field, much research is focused on fermentation of cellulosic biomass by thermophilic microbes to produce biofuels; however, little effort is dedicated to the development of imaging tools to monitor this dynamic biological process. This is, in part, due to the experimental challenges of imaging cells under both anaerobic and thermophilic conditions. Here an imaging system is described that integrates confocal microscopy, a flow cell device, and a lipophilic dye to visualize cells. Solutions to technical obstacles regarding suitable fluorescent markers, photodamage during imaging, and maintenance of environmental conditions during imaging are presented. This system was utilized to observe cellulose colonization by Clostridium thermocellum under anaerobic conditions at 60°C. This method enables live cell imaging of bacterial growth under anaerobic and thermophilic conditions and should be widely applicable to visualizing different cell types or processes in real time.

Original languageEnglish
Pages (from-to)849-856
Number of pages8
JournalJournal of Environmental Sciences (China)
Volume25
Issue number5
DOIs
StatePublished - May 1 2013

Funding

This work was supported by the BioEnergy Science Center (BESC), which is a U.S. Department of Energy Bioenergy Research Center supported by the Office of Biological and Environmental Research in the DOE Office of Science. Oak Ridge National Laboratory is managed by UT-Battelle, LLC, for the U.S. Department of Energy under contract DE-AC05-00OR22725.

Keywords

  • Biofilm
  • Biofuel
  • Cellulose
  • Confocal laser scanning microscopy
  • Lipophilic dye
  • Thermophile

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