An improved cell separation technique for marine subsurface sediments: applications for high‐throughput analysis using flow cytometry and cell sorting
-
- Yuki Morono
- Geomicrobiology Group Kochi Institute for Core Sample Research Japan Agency for Marine‐Earth Science and Technology (JAMSTEC) Monobe B200 Nankoku Kochi 783‐8502 Japan
-
- Takeshi Terada
- Marine Works Japan Ltd Oppamahigashi 3‐54‐1 Yokosuka 237‐0063 Japan
-
- Jens Kallmeyer
- Deutsches GeoForschungsZentrum GFZ Section 4.5 Geomicrobiology Telegrafenberg 14473 Potsdam Germany
-
- Fumio Inagaki
- Geomicrobiology Group Kochi Institute for Core Sample Research Japan Agency for Marine‐Earth Science and Technology (JAMSTEC) Monobe B200 Nankoku Kochi 783‐8502 Japan
書誌事項
- 公開日
- 2013-06-03
- 資源種別
- journal article
- 権利情報
-
- http://creativecommons.org/licenses/by-nc-nd/3.0/
- DOI
-
- 10.1111/1462-2920.12153
- 公開者
- Wiley
この論文をさがす
説明
<jats:title>Summary</jats:title> <jats:p> Development of an improved technique for separating microbial cells from marine sediments and standardization of a high‐throughput and discriminative cell enumeration method were conducted. We separated microbial cells from various types of marine sediment and then recovered the cells using multilayer density gradients of sodium polytungstate and/or <jats:styled-content style="fixed-case">N</jats:styled-content> ycodenz, resulting in a notably higher percent recovery of cells than previous methods. The efficiency of cell extraction generally depends on the sediment depth; using the new technique we developed, more than 80% of the total cells were recovered from shallow sediment samples (down to 100 meters in depth), whereas ∼ 50% of cells were recovered from deep samples (100–365 m in depth). The separated cells could be rapidly enumerated using flow cytometry ( <jats:styled-content style="fixed-case">FCM</jats:styled-content> ). The data were in good agreement with those obtained from manual microscopic direct counts over the range 10 <jats:sup>4</jats:sup> –10 <jats:sup>8</jats:sup> cells cm <jats:sup>−3</jats:sup> . We also demonstrated that sedimentary microbial cells can be efficiently collected using a cell sorter. The combined use of our new cell separation and <jats:styled-content style="fixed-case">FCM</jats:styled-content> /cell sorting techniques facilitates high‐throughput and precise enumeration of microbial cells in sediments and is amenable to various types of single‐cell analyses, thereby enhancing our understanding of microbial life in the largely uncharacterized deep subseafloor biosphere. </jats:p>
収録刊行物
-
- Environmental Microbiology
-
Environmental Microbiology 15 (10), 2841-2849, 2013-06-03
Wiley
