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

Title
Application of tyramide signal amplification-fluorescence in situ hybridisation and flow cytometry to detection of Heterosigma akashiwo (Raphidophyceae) in natural waters
Author
한명수
Keywords
cell cycle analysis; flow cytometer; Heterosigma akashiwo; accurate identification; tyramide signal amplification-fluorescence in situ hybridisation
Issue Date
2012-05
Publisher
Taylor & Francis
Citation
New Zealand Journal of Geology and Geophysics, 2012, 46(1), P.137-148
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.
URI
https://www.tandfonline.com/doi/abs/10.1080/00288330.2011.613404http://hdl.handle.net/20.500.11754/49893
ISSN
0028-8306; 0028-8330
DOI
10.1080/00288330.2011.613404
Appears in Collections:
COLLEGE OF NATURAL SCIENCES[S](자연과학대학) > LIFE SCIENCE(생명과학과) > Articles
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