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

J. Lee, T. Katano, M. Chang, M. S. Han

Research output: Contribution to journalArticle

3 Scopus citations

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 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.

Original languageEnglish
Pages (from-to)137-148
Number of pages12
JournalNew Zealand Journal of Marine and Freshwater Research
Volume46
Issue number1
DOIs
StatePublished - 2012 Mar 1

Keywords

  • Heterosigma akashiwo
  • accurate identification
  • cell cycle analysis
  • flow cytometer
  • tyramide signal amplification-fluorescence in situ hybridisation

Fingerprint Dive into the research topics of 'Application of tyramide signal amplification-fluorescence in situ hybridisation and flow cytometry to detection of Heterosigma akashiwo (Raphidophyceae) in natural waters'. Together they form a unique fingerprint.

  • Cite this