Simultaneous direct counting of total and specific microbial cells in seawater, using a deep-sea microbe as target |
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Authors: | Maruyama A Sunamura M |
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Institution: | Bioconsortium Analysis Program, National Institute of Bioscience and Human Technology, Higashi, Tsukuba, Ibaraki 305-8566, Japan. maruyama@nibh.go.jp |
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Abstract: | To rapidly and accurately enumerate total and specific microbes in aquatic samples, fluorescent in situ hybridization was combined with direct counting via direct immobilization of cells on a polymer-coated Nuclepore filter. The technique, named FISH-DC, achieved almost complete recovery of total cells and reproducibility of Psychrobacter pacificensis cells of deep-sea origin (error, =3%) in a mixed culture and in natural seawater. Target cells immobilized on the filter were also successfully enumerated after stringent 3-cycle hybridization and even after a 16-month preservation at -30 degrees C. |
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