Application of tyramide signal amplification-fluorescence in situ hybridisation and flow cytometry to detection of Heterosigma akashiwo (Raphidophyceae) in natural waters SCIE SCOPUS

Cited 4 time in WEB OF SCIENCE Cited 4 time in Scopus
Title
Application of tyramide signal amplification-fluorescence in situ hybridisation and flow cytometry to detection of Heterosigma akashiwo (Raphidophyceae) in natural waters
Author(s)
Lee, J.; Katano, T.; Chang, M.; Han, M-S
Publication Year
2012-03
Abstract
We developed protocols to analyse the cell cycle of Heterosigma akashiwo in the natural phytoplankton assemblage using tyramide signal amplification-fluorescence in situ hybridisation (TSA-FISH) and flow cytometry. We determined the optimum probe and formamide concentrations for the species-specific TSA-FISH probe to be 0.5 ng mu L-1 and 40%, respectively. The probe did not hybridise to non-target phytoplankton including Chattonella sp., which is closely related to Heterosigma and has high sequence similarity to it. We could successfully identify G1- and G2-phase cells and the diel cell cycle of H. akashiwo not only in the laboratory but also in the field after TSA-FISH. These results demonstrate that TSA-FISH for H. akashiwo in natural coastal waters could serve as a powerful tool for understanding their bloom mechanism using flow cytometry.
ISSN
0028-8330
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/3617
DOI
10.1080/00288330.2011.613404
Bibliographic Citation
NEW ZEALAND JOURNAL OF MARINE AND FRESHWATER RESEARCH, v.46, no.1, pp.137 - 148, 2012
Publisher
TAYLOR & FRANCIS LTD
Subject
CELL-CYCLE REGULATION; TARGETED OLIGONUCLEOTIDE PROBES; RIBOSOMAL-RNA; MARINE SYNECHOCOCCUS; RED TIDE; DIEL PATTERNS; GROWTH-RATES; NEW-ZEALAND; TOKYO BAY; IDENTIFICATION
Keywords
cell cycle analysis; flow cytometer; Heterosigma akashiwo; accurate identification; tyramide signal amplification-fluorescence in situ hybridisation
Type
Article
Language
English
Document Type
Article
Publisher
TAYLOR & FRANCIS LTD
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