Related Experiment Video
Updated: Oct 1, 2025
Simultaneous Imaging and Flow-Cytometry-based Detection of Multiple Fluorescent Senescence Markers in Therapy-Induced Senescent Cancer Cells
Published on: July 12, 2022
Flow cytometric single cell-based assay to simultaneously detect cell death, cell cycling, DNA content and cell
Elizabeth Lieschke1,2, Zilu Wang1,2, Catherine Chang1
1The Walter and Eliza Hall Institute of Medical Research, Melbourne, VIC, Australia.
Abstract:
Cell death, cell cycle arrest and cellular senescence are three distinct cellular responses that can be induced by oncogene activation and diverse anti-cancer agents, and this often requires the action of the tumour suppressor TP53. Within a cell population, or even within an individual cell, these processes are not necessarily mutually exclusive. It is therefore important to measure all these processes simultaneously. However, current assays generally visualise only one or at best two responses, often only detecting the dominant one. Here, we present a novel flow cytometric assay that allows simultaneous assessment of cell viability and cell cycling through measurement of DNA content and DNA synthesis, and markers of cell senescence at the single cell level. We demonstrate that this assay can be performed on both human and murine cells, that are either cancerous or non-transformed, and can help to dissect complex cell fate decisions. We believe that this experimental tool will be useful for the study of diverse biological processes.
Insights
This study introduces a novel flow cytometry assay for simultaneously measuring cell death, cell cycle arrest, and cellular senescence. This tool aids in dissecting complex cell fate decisions in various cell types.
Area of Science:
- Cellular biology
- Cancer research
- Molecular oncology
Background:
- Cell death, cell cycle arrest, and cellular senescence are critical responses to oncogene activation and anti-cancer agents, often involving the tumor suppressor TP53.
- These cellular processes are not mutually exclusive and can occur concurrently within a cell population or an individual cell.
- Current assays are limited, often visualizing only one or two responses and potentially missing the dominant outcome.
Purpose of the Study:
- To develop a novel flow cytometric assay for simultaneous assessment of multiple cellular responses.
- To enable single-cell level analysis of cell viability, cell cycling, and senescence.
- To provide a tool for dissecting complex cell fate decisions.
Main Methods:
- Development of a novel flow cytometry assay.
- Simultaneous measurement of DNA content (cell cycling) and DNA synthesis.
- Assessment of senescence markers at the single-cell level.
Main Results:
- The assay successfully allows for the simultaneous assessment of cell viability, cell cycling, and senescence.
- Demonstrated applicability to both human and murine cells, including cancerous and non-transformed cell lines.
- The assay effectively dissects complex cell fate decisions.
Conclusions:
- The developed flow cytometry assay is a valuable experimental tool for biological research.
- Enables comprehensive analysis of cellular responses, overcoming limitations of existing methods.
- Facilitates the study of diverse biological processes, particularly in cancer research and drug development.
