title: Label free high throughput screening for apoptosis inducing chemicals using time-lapse microscopy signal processing.
abstract: Label free time-lapse microscopy has opened a new avenue to the study of time evolving events in living cells. When combined with automated image analysis it provides a powerful tool that enables automated large-scale spatiotemporal quantification at the cell population level. Very few attempts, however, have been reported regarding the design of image analysis algorithms dedicated to the detection of apoptotic cells in such time-lapse microscopy images. In particular, none of the reported attempts is based on sufficiently fast signal processing algorithms to enable large-scale detection of apoptosis within hours/days without access to high-end computers. Here we show that it is indeed possible to successfully detect chemically induced apoptosis by applying a two-dimensional linear matched filter tailored to the detection of objects with the typical features of an apoptotic cell in phase-contrast images. First a set of recorded computational detections of apoptosis was validated by comparison with apoptosis specific caspase activity readouts obtained via a fluorescence based assay. Then a large screen encompassing 2,866 drug like compounds was performed using the human colorectal carcinoma cell line HCT116. In addition to many well known inducers (positive controls) the screening resulted in the detection of two compounds here reported for the first time to induce apoptosis.
authors.affiliation: Department of Medical Sciences, Cancer Pharmacology and Computational Medicine, Uppsala Academic Hospital, Uppsala University, 751 85, Uppsala, Sweden.
authors.forename: Obaid
authors.initials: O
authors.lastname: Aftab
authors.forename: Madiha
authors.initials: M
authors.lastname: Nazir
authors.forename: Mårten
authors.initials: M
authors.lastname: Fryknäs
authors.forename: Ulf
authors.initials: U
authors.lastname: Hammerling
authors.forename: Rolf
authors.initials: R
authors.lastname: Larsson
authors.forename: Mats G
authors.initials: MG
authors.lastname: Gustafsson
mesh_terms: D000903:Antibiotics, Antineoplastic / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism; D053179:Caspase 7 / Q000378:metabolism; D045325:HCT116 Cells; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D008853:Microscopy; D016685:Mitomycin / Q000494:pharmacology; D009285:Naphthoquinones / Q000494:pharmacology; D009930:Organic Chemicals / Q000494:pharmacology*; D010880:Piperidines / Q000494:pharmacology; D013194:Staining and Labeling / Q000379:methods; D059008:Time-Lapse Imaging
chemical_list: D000903:Antibiotics, Antineoplastic; D009285:Naphthoquinones; D009930:Organic Chemicals; D010880:Piperidines; C027013:diperodon; D016685:Mitomycin; C063002:2,3-dimethoxy-1,4-naphthoquinone; D053148:Caspase 3; D053179:Caspase 7