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Cellular imaging assay for early evaluation of an apoptosis inducer

Apoptosis volume 16, Article number: 1068 (2011) Cite this article

Abstract

The objective of this study was to propose a feasibility of a cellular imaging assay as an alternative to the conventional cytotoxicity assay, such as MTS assay, for apoptosis monitoring. As an apoptosis monitoring parameter, affinity interaction between phosphatidylserine (PS) and annexin V was chosen. First, the specific binding affinity between annexin V and PS in phospholipid bilayers consisting of various molar (0–15%) composition of PS was measured using a surface plasmon resonance biosensor. As PS composition increased, the binding level of annexin V increased proportionally. Second, various concentrations (0.1–10 μM) of staurosporine were used as to induce apoptosis and introduced to MCF-7 breast carcinoma cells. The cellular fluorescence images from annexin V-FITC conjugate were obtained by confocal microscopy, and their fluorescence intensities were quantified by image scanning. Dose–apoptosis (or cell death) relationships were very similar to those from MTS and FACS assays. In summary, our cellular imaging method could serve as a quicker and simpler alternative to MTS (end point assay) and FACS (flow cytometry) to screen potential apoptosis inducers.

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Acknowledgments

This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (R11-2008-044-01001-0). It was also supported by Kyungwon University Research Fund in 2010.

Competing interest

The authors declare no competing interests.

Author information

Author notes

  1. Sang Youn Hwang

    Present address: Hanmi Pharmaceutics Research Center, Giheung, Korea

  2. Seung Hun Cho

    Present address: Korean Research Institute of Biosciences and Biotechnology, Daejeon, Korea

Affiliations

  1. Department of Chemical Engineering, Hanyang University, Ansan, Korea

    Sang Youn Hwang

  2. Department of Bionanotechnology, Hanyang University, Ansan, Korea

    Seung Hun Cho

  3. Department of Biochemistry and Cell Biology School of Medicine, Kyungpook National University, Daegu, Korea

    Byung Heon Lee

  4. Department of Life Science, Kyungwon University, Seongnam, Korea

    Yoon-Jae Song

  5. College of Bionanotechnology, Kyungwon University, Seongnam, Korea

    E. K. Lee

Authors
  1. Sang Youn Hwang
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  2. Seung Hun Cho
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  3. Byung Heon Lee
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Correspondence to E. K. Lee.

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Hwang, S.Y., Cho, S.H., Lee, B.H. et al. Cellular imaging assay for early evaluation of an apoptosis inducer. Apoptosis 16, 1068 (2011). https://doi.org/10.1007/s10495-011-0636-7

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  • DOI: https://doi.org/10.1007/s10495-011-0636-7

Keywords

  • Apoptosis
  • Cell death
  • Phosphatidylserine
  • Annexin V
  • Staurosporine
  • Confocal microscopy
  • MTS assay
  • Early screening