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Apoptosis detection via automated algorithms to analyze biomarker translocation in reporter cells
Author(s) -
Ahmed A. H. Rezwanuddin,
DereliKorkut Zeynep,
Lee Joanne Haeun,
Piracha Sidra,
Gilchrist M. Lane,
Jiang Xuejun,
Wang Sihong
Publication year - 2020
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.27280
Subject(s) - computer science , algorithm , workflow , apoptosis , throughput , reporter gene , computational biology , chemistry , biology , biochemistry , gene , telecommunications , gene expression , database , wireless
Abstract Rapid, efficient, and robust quantitative analyses of dynamic apoptotic events are essential in a high‐throughput screening workflow. Currently used methods have several bottlenecks, specifically, limitations in available fluorophores for downstream assays and misinterpretation of statistical image data analysis. In this study, we developed cytochrome‐C (Cyt‐C) and caspase‐3/‐8 reporter cell lines using lung (PC9) and breast (T47D) cancer cells, and characterized them from the response to apoptotic stimuli. In these two reporter cell lines, the spatial fluorescent signal translocation patterns served as reporters of activations of apoptotic events, such as Cyt‐C release and caspase‐3/‐8 activation. We also developed a vision‐based, tunable, automated algorithm in MATLAB to implement the robust and accurate analysis of signal translocation in single or multiple cells. Construction of the reporter cell lines allows live monitoring of apoptotic events without the need for any other dyes or fixatives. Our algorithmic implementation forgoes the use of simple image statistics for more robust analytics. Our optimized algorithm can achieve a precision greater than 90% and a sensitivity higher than 85%. Combining our automated algorithm with reporter cells bearing a single‐color dye/fluorophore, we expect our approach to become an integral component in the high‐throughput drug screening workflow.