Open AccessSimultaneous Direct Counting of Total and Specific Microbial Cells in Seawater, Using a Deep-Sea Microbe as Target
Author(s) -
Akihiko Maruyama,
Michinari Sunamura
Publication year - 2000
Publication title -
applied and environmental microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.552
H-Index - 324
eISSN - 1070-6291
pISSN - 0099-2240
DOI - 10.1128/aem.66.5.2211-2215.2000
Subject(s) - seawater , reproducibility , membrane filter , fish <actinopterygii> , filter (signal processing) , chromatography , biology , in situ , fluorescence , fluorescence in situ hybridization , chemistry , membrane , ecology , biochemistry , fishery , gene , physics , organic chemistry , chromosome , quantum mechanics , computer science , computer vision
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 ofPsychrobacter 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°C.