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Flow Cytometric Analysis of Apoptotic Biomarkers in Actinomycin D-Treated SiHa Cervical Cancer Cells
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SYTO probes: markers of apoptotic cell demise

Donald Wlodkowic1, Joanna Skommer

  • 1University of Glasgow, UK.

Current Protocols in Cytometry
|September 5, 2008
PubMed

Insights

New SYTO probes offer a rapid, high-throughput method for detecting apoptosis, a key type of tumor cell death. These DNA-binding dyes are valuable tools for multiparameter flow cytometry in cancer research.

Area of Science:

  • Biotechnology
  • Molecular Biology
  • Cancer Research

Background:

  • Advancing mechanistic studies of tumor cell death requires efficient analytical tools.
  • SYTO probes are emerging as user-friendly markers for caspase-dependent apoptosis.
  • Their spectral properties facilitate multiparameter flow cytometry assay development.

Purpose of the Study:

  • To provide protocols for multiparametric assessment of apoptotic events using SYTO probes.
  • To demonstrate the utility of SYTO probes in conjunction with other cell death markers.

Main Methods:

  • Utilizing green fluorescent SYTO 16 and red fluorescent SYTO 17 dyes with plasma membrane permeability markers.
  • Employing SYTO 16 with tetramethylrhodamine methyl ester (TMRM) to assess mitochondrial membrane potential.
  • Combining SYTO 16 with 7-aminoactinomycin D (7-AAD) as a plasma membrane permeability marker.

Main Results:

  • Established protocols for multiparametric apoptosis detection using SYTO probes.
  • Demonstrated successful combination of SYTO probes with plasma membrane and mitochondrial potential markers.
  • Showcased the versatility of SYTO probes in diverse cell death assays.

Conclusions:

  • SYTO probes are effective for high-throughput, multiparametric analysis of apoptotic cell death.
  • These probes are valuable for tracking apoptosis in various cancer cell lines and primary samples.
  • The provided protocols enable advanced flow cytometry assays for cancer research.

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