title: Simultaneous direct counting of total and specific microbial cells in seawater, using a deep-sea microbe as target.
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.
authors.affiliation: Bioconsortium Analysis Program, National Institute of Bioscience and Human Technology, Higashi, Tsukuba, Ibaraki 305-8566, Japan. maruyama@nibh.go.jp
authors.forename: A
authors.initials: A
authors.lastname: Maruyama
authors.forename: M
authors.initials: M
authors.lastname: Sunamura
mesh_terms: D001407:Bacillus / Q000166:cytology / Q000302:isolation & purification; D001431:Bacteriological Techniques; D018914:Cells, Immobilized; D020563:Gammaproteobacteria / Q000166:cytology* / Q000302:isolation & purification*; D017404:In Situ Hybridization, Fluorescence; D011550:Pseudomonas aeruginosa / Q000166:cytology / Q000302:isolation & purification; D015203:Reproducibility of Results; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000302:isolation & purification; D012623:Seawater / Q000382:microbiology*; D014733:Vibrio / Q000166:cytology / Q000302:isolation & purification