Automatic image analysis for quantification of apoptosis in animal cell culture by annexin-V affinity assay

Pläsier, Birgitt; Lloyd, D. R.; Paul, Goutam; Thomas, C. R.; Al‐Rubeai, Mohamed

doi:10.1016/s0022-1759(99)00107-6

Cited by 39 publications

(34 citation statements)

References 34 publications

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“…1 A and B). There are multiple approaches for extracting phenotypic information, including detecting either known biological phenotypes [e.g., apoptosis (31) or cell cycle phases (32) or general image properties, e.g., cell morphology and patterns of marker intensities (14,18)]. Here, we found that a feature set designed to measure the degree of marker colocalization performed well in capturing phenotypic differences in signaling state among visually distinct cells.…”

Section: Resultsmentioning

confidence: 86%

Characterizing heterogeneous cellular responses to perturbations

Slack

Martínez

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

152

175

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Cellular populations have been widely observed to respond heterogeneously to perturbation. However, interpreting the observed heterogeneity is an extremely challenging problem because of the complexity of possible cellular phenotypes, the large dimension of potential perturbations, and the lack of methods for separating meaningful biological information from noise. Here, we develop an image-based approach to characterize cellular phenotypes based on patterns of signaling marker colocalization. Heterogeneous cellular populations are characterized as mixtures of phenotypically distinct subpopulations, and responses to perturbations are summarized succinctly as probabilistic redistributions of these mixtures. We apply our method to characterize the heterogeneous responses of cancer cells to a panel of drugs. We find that cells treated with drugs of (dis-)similar mechanism exhibit (dis-)similar patterns of heterogeneity. Despite the observed phenotypic diversity of cells observed within our data, low-complexity models of heterogeneity were sufficient to distinguish most classes of drug mechanism. Our approach offers a computational framework for assessing the complexity of cellular heterogeneity, investigating the degree to which perturbations induce redistributions of a limited, but nontrivial, repertoire of underlying states and revealing functional significance contained within distinct patterns of heterogeneous responses. automated microscopy ͉ cellular heterogeneity ͉ image analysis

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Section: Resultsmentioning

confidence: 86%

Characterizing heterogeneous cellular responses to perturbations

Slack

Martínez

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

152

175

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“…Annexin V, a calcium dependent phospholipidbinding protein with a high affinity for PS, can therefore be used as a sensitive probe for the exposure of PS on the cell membrane and hence as a marker of apoptosis. 22) Annexin V+ and 7-AAD− cells were designated as early apoptotic and Annnexin V+ and 7-AAD+ cells were designated as necrotic. More apoptotic cells were observed in 20(S)-GRh2 treated group (20,40, 50 µM) than in control group (Figs.…”

Section: Effect Of 20(s)-grh2 On Nuclear Morphology Of Reh Cellsmentioning

confidence: 99%

20(<i>S</i>)-Ginsenoside Rh2 Induces Apoptosis in Human Leukaemia Reh Cells through Mitochondrial Signaling Pathways

Xia

Wang

et al. 2014

Biological & Pharmaceutical Bulletin

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“…1 The clearest assessment of cell apoptosis is gained from microscopy or flow cytometry analyses that examine large cell populations and identify and quantify subpopulations. A common method of detecting PS on a cell surface is to use the PS-binding protein annexin V. 2,3 This 35 kDa protein is typically labeled with a fluorescent dye, often fluorescein because it can be conveniently excited by an argon laser. Although annexin V is a useful reagent, it is an expensive protein, and there are some technical drawbacks.…”

Section: Dear Editormentioning

confidence: 99%

“…Not only should PSS-380 be useful as an apoptosis sensor in cell biology research, it may also have utility in high-throughput assays for drug discovery, as well as automated monitoring of animal cell culture technology. 3 A potential technical drawback with PSS-380 is that it requires UV excitation, a feature that is presently not available on common flow cytometers. Our current research efforts are focused on making analogues of PSS-380 that have altered fluorescence properties.…”

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confidence: 99%