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Updated: Aug 17, 2026
Flow Cytometric Analysis of Apoptotic Biomarkers in Actinomycin D-Treated SiHa Cervical Cancer Cells
Published on: August 26, 2021
Screening for apoptosis--classical and emerging techniques
Henrik Lövborg1, Joachim Gullbo, Rolf Larsson
1Department of Medical Sciences, Division of Clinical Pharmacology, University Hospital, Uppsala University, Sweden. henrik.lovborg@medsci.uu.se
Abstract:
There has been a rapid development of cell-based assays and screening methods to identify promising apoptosis-inducing drug candidates for the treatment of cancer. Distinguishing between the complex processes involved in apoptosis and other forms of cell death requires information on both biochemical and morphological processes in the cell. Traditionally, many assays have been limited to measuring, for example, caspase activity using fluorogenic substrates. However, these screening assays provide only limited information on the complex processes involved in apoptosis. In this review we describe some of the available apoptosis assays amenable to high-throughput screening. In particular, image-based high-content screening assays to evaluate multiple biochemical and morphological parameters in apoptotic cells are described. Through combining the imaging of cells in microtiter plates with powerful image analysis algorithms, one can acquire deeper knowledge on multiple biochemical or morphological pathways at the single-cell level at an early stage in the development of novel anti-cancer drugs.
Insights
High-throughput screening assays are crucial for identifying cancer drug candidates that induce apoptosis. Image-based high-content screening offers deeper insights into cellular processes for effective drug development.
Area of Science:
- Oncology
- Cell Biology
- Drug Discovery
Background:
- Cell-based assays are rapidly advancing for identifying apoptosis-inducing cancer drug candidates.
- Traditional assays, like caspase activity measurement, offer limited insight into complex cell death processes.
- Distinguishing apoptosis from other cell death requires biochemical and morphological data.
Purpose of the Study:
- To review apoptosis assays suitable for high-throughput screening (HTS).
- To highlight image-based high-content screening (HCS) for evaluating multiple apoptotic parameters.
- To emphasize the utility of HCS in early-stage anti-cancer drug development.
Main Methods:
- Review of existing literature on apoptosis assays for HTS.
- Description of image-based HCS methodologies.
- Integration of microtiter plate imaging with advanced image analysis algorithms.
Main Results:
- Identified various apoptosis assays compatible with HTS.
- Detailed the application of image-based HCS for multi-parametric analysis of apoptotic cells.
- Demonstrated the potential for acquiring single-cell level data on biochemical and morphological pathways.
Conclusions:
- Image-based HCS provides comprehensive data on apoptosis.
- This approach enhances understanding of cellular pathways in drug development.
- HCS is valuable for early-stage identification of novel anti-cancer drugs.
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