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Updated: May 10, 2026
Live-cell Imaging of Lysosomal Membrane Permeabilization During Necroptosis
Published on: November 14, 2025
Time-lapse imaging of necrosis
Fredrik Wållberg1, Tencho Tenev, Pascal Meier
1Chester Beatty Laboratories, The Breakthrough Toby Robins Breast Cancer Research Centre, Institute of Cancer Research, London, UK.
Abstract:
The processes of dying are as tightly regulated as those of growth and proliferation. Recent work into the molecular pathways that regulate and execute cell death have uncovered a plethora of signalling cascades that lead to distinct modes of cell death, including "apoptosis," "necrosis," "autophagic cell death," and "mitotic catastrophe." Given that cells can readily switch from one form of death to another, it is vital to carefully monitor the form of death under investigation. Particularly, end-point techniques are intrinsically unsuitable for assessing apoptosis versus necrosis, as they cannot reconstruct the sequence of events that have led to cell death. Since apoptotic cells frequently undergo secondary necrosis under in vitro culture conditions, novel methods relying on high-throughput time-lapse fluorescence video microscopy have been developed. Here we describe the use of this technique to reliably distinguish necrosis from apoptosis and secondary necrosis.
Insights
Understanding cell death is crucial. This study introduces a novel microscopy technique to accurately differentiate apoptosis, necrosis, and secondary necrosis, overcoming limitations of traditional methods.
Area of Science:
- Cell Biology
- Molecular Biology
- Biotechnology
Background:
- Cell death is a highly regulated biological process, similar to growth and proliferation.
- Multiple distinct modes of cell death exist, including apoptosis, necrosis, autophagic cell death, and mitotic catastrophe.
- Distinguishing between these cell death modalities is challenging due to the dynamic and interconnected nature of cell death pathways.
Purpose of the Study:
- To address the limitations of end-point techniques in distinguishing between apoptosis and necrosis.
- To develop and validate a method for reliably differentiating various forms of cell death, including secondary necrosis.
- To provide a tool for accurate assessment of cell death mechanisms in biological research.
Main Methods:
- Utilized high-throughput time-lapse fluorescence video microscopy.
- Developed novel methods to monitor cellular events over time.
- Applied techniques to distinguish apoptosis, necrosis, and secondary necrosis in real-time.
Main Results:
- Successfully differentiated apoptosis from necrosis and secondary necrosis using time-lapse microscopy.
- Demonstrated the inadequacy of end-point assays for precise cell death modality assessment.
- Showcased the ability of the technique to capture the sequential events leading to cell death.
Conclusions:
- High-throughput time-lapse fluorescence video microscopy is a powerful tool for distinguishing cell death modalities.
- This technique overcomes the limitations of static assays in studying dynamic cell death processes.
- Accurate discrimination of apoptosis, necrosis, and secondary necrosis is vital for understanding cell fate and disease.
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