Apoptosis detection via automated algorithms to analyze biomarker translocation in reporter cells Journal Article


Authors: Ahmed, A. H. R.; Dereli-Korkut, Z.; Lee, J. H.; Piracha, S.; Gilchrist, M. L.; Jiang, X.; Wang, S.
Article Title: Apoptosis detection via automated algorithms to analyze biomarker translocation in reporter cells
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. © 2020 Wiley Periodicals, Inc.
Keywords: controlled study; human cell; antineoplastic agent; biomarkers; cell death; apoptosis; confocal microscopy; image analysis; cell line; caspase 3; enzyme activation; high throughput screening; caspases; automation; amino acid sequence; cell culture; cancer cell; tumor necrosis factor related apoptosis inducing ligand; protein secretion; cellular distribution; caspase 8; dye; nuclear localization signal; fluorescence activated cell sorting; mitochondrion; chemical activation; cytochrome c; detection algorithm; cytochrome-c; caspase-8; automated algorithms; time lapse imaging; reporter cell line; nuclear export signal; human; article; caspase-3; t-47d cell line; pc-9 cell line; translocation pattern; biomarker translocation; reporter cells; optimized algorithms; signal analysis
Journal Title: Biotechnology and Bioengineering
Volume: 117
Issue: 5
ISSN: 0006-3592
Publisher: Wiley Blackwell  
Date Published: 2020-05-01
Start Page: 1470
End Page: 1482
Language: English
DOI: 10.1002/bit.27280
PUBMED: 31956989
PROVIDER: scopus
DOI/URL:
Notes: Article -- Source: Scopus
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  1. Xuejun Jiang
    92 Jiang