An improved method for high throughput discrimination and enumeration of sedimentary cells using flow cytometry

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Author(s): Morono, Y.; Kallmeyer, J.; Terada, T.; Inagaki, F.
Integrated Ocean Drilling Program, Expedition 329, Shipboard Scientific Party
Author Affiliation(s): Primary:
Japan Agency of Marine-Earth Science and Technology, Kochi Institute for Core Sample Research, Kochi, Japan
Other:
University of Potsdam, Germany
Marine Works Japan, Japan
Volume Title: AGU 2011 fall meeting
Source: American Geophysical Union Fall Meeting, Vol.2011; American Geophysical Union 2011 fall meeting, San Francisco, CA, Dec. 5-9, 2011. Publisher: American Geophysical Union, Washington, DC, United States
Note: In English
Summary: Detection and enumeration of microbial life in marine subsurface environments provides primary information on the extent and habitability of the Earth's biosphere. Flow cytometry (FCM) is a powerful tool for identifying and enumerating fluorescence-stained cells with high throughput, using fluorescent intensity, range of wavelength, and cell size. FCM is widely used in medical sciences and aquatic microbial ecology. However, mineral grains and difficulties in distinguishing between life cells and non-specific background fluorescence prevented FCM to be applied for counting microbial cells in sediment or rock samples. SYBR Green I-stained cells can be distinguished from non-biological background signals based on differences in their fluorescence spectra. Here we extended this technique to FCM analysis by modifying the cell detachment protocol using a density gradient method, and then standardized an FCM cell counting method for various types of marine subsurface sediments. Microbial cells in sediment samples could effectively be detached and analyzed discriminatively with FCM. The high capacity of FCM to count particles (up to 10,000 cells/sec) and its high sensitivity will provide information about microbial cell abundance at high spatial resolution and with unprecedented accuracy. This improved cell count method will be useful to evaluate samples with high depth resolution, including narrow geochemical and geological interfaces as potential specific microbial niches, and may even help to assess very low population densities at the fringe of the biosphere.
Year of Publication: 2011
Research Program: IODP Integrated Ocean Drilling Program
Key Words: 22 Environmental Geology; Bedrock; Biosphere; Cytometry; East Pacific; Ecology; Ecosystems; Expedition 329; Fluorescence; Habitat; Integrated Ocean Drilling Program; Microorganisms; Pacific Ocean; Sediments; South Pacific; Southeast Pacific; Underground space
Coordinates: S455800 S235100 W1230900 W1660000
Record ID: 2014024643
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