title: Cellular imaging assay for early evaluation of an apoptosis inducer.
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.
authors.affiliation: Department of Chemical Engineering, Hanyang University, Ansan, Korea.
authors.forename: Sang Youn
authors.initials: SY
authors.lastname: Hwang
authors.forename: Seung Hun
authors.initials: SH
authors.lastname: Cho
authors.forename: Byung Heon
authors.initials: BH
authors.lastname: Lee
authors.forename: Yoon-Jae
authors.initials: YJ
authors.lastname: Song
authors.forename: E K
authors.initials: EK
authors.lastname: Lee
mesh_terms: D017304:Annexin A5 / Q000378:metabolism*; D017209:Apoptosis / Q000187:drug effects*; D001943:Breast Neoplasms / Q000473:pathology; D045744:Cell Line, Tumor; D003584:Cytological Techniques / Q000379:methods*; D005260:Female; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D006801:Humans; D018613:Microscopy, Confocal; D010718:Phosphatidylserines / Q000378:metabolism*; D019311:Staurosporine / Q000494:pharmacology; D020349:Surface Plasmon Resonance; D013778:Tetrazolium Salts; D013844:Thiazoles
chemical_list: D017304:Annexin A5; D005456:Fluorescent Dyes; D010718:Phosphatidylserines; D013778:Tetrazolium Salts; D013844:Thiazoles; C070380:3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium; D019311:Staurosporine; D016650:Fluorescein-5-isothiocyanate