Applications of immuno-magnetic bead and immunofluorescent flow cytometric techniques for the quantitative detection of HAB microalgae

Jian Huang; Ruobing Wen; Zhenmin Bao; Zhenghong Sui; Ningbo Sun; Kyoungho Kang

doi:10.1007/s00343-012-1208-6

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Applications of immuno-magnetic bead and immunofluorescent flow cytometric techniques for the quantitative detection of HAB microalgae

Updated：2023-05-16

- Applications of immuno-magnetic bead and immunofluorescent flow cytometric techniques for the quantitative detection of HAB microalgae
- Journal of Oceanology and Limnology Vol. 30, Issue 3, Pages: 433-439(2012)
- Affiliations：
  
  1. Key Laboratory of Marine Genetics and Breeding of Ministry of Education Ocean University of China
  2. North China Sea Environmental Monitoring Center State Oceanic Administration
  3. The Fifth Middle School of Zibo
  4. Department of Aquaculture Chonnam National University
- Author bio：
- Funds：
- DOI：10.1007/s00343-012-1208-6
  CLC：
- Online First：12 December 2011，
  
  Published：2012
- Accepted：
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Huang, J., Wen, R., Bao, Z. et al. Applications of immuno-magnetic bead and immunofluorescent flow cytometric techniques for the quantitative detection of HAB microalgae., Chin. J. Ocean. Limnol. 30, 433–439 (2012). https://doi.org/10.1007/s00343-012-1208-6
DOI：

Huang, J., Wen, R., Bao, Z. et al. Applications of immuno-magnetic bead and immunofluorescent flow cytometric techniques for the quantitative detection of HAB microalgae., Chin. J. Ocean. Limnol. 30, 433–439 (2012). https://doi.org/10.1007/s00343-012-1208-6 DOI：

摘要

Abstract

Over the last several decades

harmful algal blooms (HABs) have become a serious environmental problem in many parts of the world. A rapid and accurate detection process for HAB algae has yet to be developed. Heterosigma akashiwo is one of the most important HABs species in China. The objective of this study was to develop an immunologic technique that can rapidly and sensitively count H. akashiwo cells. Five HABs species (Alexandrium catenella

Thalassiosira sp.

Cryptomonas sp.

Alexandrium tamarense and Symbiodinium sp.)

were used in this study to evaluate the analysis process we developed. A polyclonal antibody with high titers against H. akashiwo was obtained by injecting H. akashiwo cells into rabbits. Immuno-magnetic beads (IMB) were produced via conjugated polyclonal antibodies with magnetic beads and applied to isolate and count H. akashiwo cells from the culture. Results show that 66.7%–91.6% of the cells were captured from unialgal culture by IMBs

and only 5.3%–12.5% of the four other HAB microalgae species were captured

indicating that the constructed IMBs combined specifically with the H. akashiwo cells. At the same time

flow cytometry (FCM) sorting was exploited to screen H. akashiwo cells after labeling with FITC conjugated polyclonal antibodies. Using the FCM technique

91.7% of the targeted cells were sorted out from mixed microalgae samples in just a few minutes. These results indicate that both antibody-involved IMB and antibody-based FCM techniques are highly effective at detecting and quantifying HAB species. These techniques

especially immuno-magnetic separation

have low associated cost

and are fast and simple processes compared with other techniques currently in use.

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Keywords

references

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