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Quantitative Detection of Apoptotic Cells Using Annexin V and Phosphatidylserine Binding by Confocal Microscopy

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At present, cell-based drug screening assays have been attracted attention for various molecular interactions including induction of apoptosis. Apoptosis is the process of programmed cell death through a tightly controlled program that plays important roles in many normal processes, ranging from fetal development to adult tissue homeostasis.1) Screening of apoptosis would allow
both early detection of therapy efficiency and evaluation of disease progression.2) In this study, the affinity of specific binding between annexin V and liposome that consists of several molar ratio of phosphatidylserine (PS) and phosphatidylcholine (PC) in presence of Ca2+ was measured by surface plasmon resonance (SPR) analysis. Annexin V-FITC is used for quantification to confirm efficacy of anticancer agent. Induction of apoptosis is induced by
stausrosporine (SSP) as one of anticancer agents. A variety of concentration of SSP induced to be apoptotic cells with macrophage cell line (RAW 264.7 cell) and breast cancer cell line (MCF-7 cell). The apoptotic, necrotic and live cells was monitored using fluorescent probes such as annexin V-FITC and propidium iodide (PI) by confocal microscopy. The fluorescent imaging data
based on confocal microscopy was analysed with intensity of fluorescence for quantification of apoptotic cells.

저자정보

  • Seung Hun Cho Dept. of Bionanotechnology
  • Do Young Cho Dept. of Chemical Engineering, Hanyang University
  • Sang Youn Hwang Dept. of Chemical Engineering, Hanyang University
  • Eun Kyu Lee Dept. of Bionanotechnology, Dept. of Chemical Engineering, Hanyang University

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