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Related Experiment Video

Updated: Jul 4, 2025

Use of a Caspase Multiplexing Assay to Determine Apoptosis in a Hypothalamic Cell Model
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A Multiplex Assay to Simultaneously Monitor Apoptosis and Necrosis Using the Cellaca® PLX Image Cytometer.

Mackenzie Pierce1, Yongyang Huang1, Allen Lin1

  • 1Department of Advanced Technology R&D, Revvity Health Sciences, Inc., 360 Merrimack St., Suite 200, Lawrence, MA, 01843, USA.

Journal of Fluorescence
|January 31, 2024
PubMed
Summary

This study presents a new multiplex detection method to monitor programmed cell death (apoptosis) and necrosis in cancer cells. The method effectively tracks apoptosis progression and aids in screening potential cancer therapeutics.

Keywords:
AnnexinApoptosisCaspaseCellaca® PLXImage cytometryNecrosis

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Area of Science:

  • Cell Biology
  • Cancer Research
  • Drug Discovery

Background:

  • Apoptosis is a crucial programmed cell death pathway for homeostasis.
  • Cancer cells often evade apoptosis, contributing to their survival and proliferation.
  • Characterizing therapeutics that induce apoptosis is vital for drug discovery.

Purpose of the Study:

  • To develop and demonstrate a multiplex detection method for monitoring apoptosis and necrosis.
  • To utilize image cytometry for simultaneous assessment of early-stage apoptosis, late-stage apoptosis, and necrosis.
  • To evaluate the method's efficacy in characterizing cancer therapeutic candidates.

Main Methods:

  • Developed a multiplex detection assay using Annexin V, Caspase-3, and Propidium Iodide (PI).
  • Employed the Cellaca® PLX Image Cytometer for apoptosis and necrosis monitoring.
  • Induced apoptosis in Jurkat and K562 cell lines with staurosporine and assessed at multiple timepoints.

Main Results:

  • Successfully detected apoptotic factors and cascades from early- to late-stage apoptosis and necrosis.
  • Observed distinct trends in apoptotic and necrotic populations within the first 1.5 hours in Jurkat cells.
  • K562 cells showed limited response to staurosporine, indicating differential sensitivity.

Conclusions:

  • The developed image cytometric method enables efficient and reliable simultaneous detection of apoptosis and necrosis.
  • This approach can significantly aid researchers in characterizing and screening potential cancer therapeutics.
  • The method offers a higher throughput solution for evaluating drug efficacy in cancer treatment.