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Describe the adaptation of flow cytometric apoptosis assays to laser scanning cytometry and demonstrate the value of combining quantitative fluorescence with visual morphological confirmation.

March 1, 1999Cytometry

Analysis of apoptosis by laser scanning cytometry

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EBElżbieta BednerCancer Research Institute XLXun LiGuangxi University WGWojciech GorczycaRejonowe Przedsiębiorstwo Wodociągów i Kanalizacji W Sosnowcu (Poland)

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Implication

Methodological review demonstrates multiparametric apoptosis detection in cell models, highlighting how laser scanning cytometry couples fluorescence measurements with visual morphology.

Key Points

Describe the adaptation of flow cytometric apoptosis assays to laser scanning cytometry and demonstrate the value of combining quantitative fluorescence with visual morphological confirmation.
Adapted flow cytometry assays—including mitochondrial membrane potential probes, annexin V, and DNA fragmentation analyses—to slide-based laser scanning cytometry.
Quantified chromatin condensation using maximal pixel fluorescence values of DNA-bound fluorochromes on immobilized cells.
Correlated fluorescence profiles with visual microscopic morphology to distinguish true apoptotic cells from phagocytic macrophages containing engulfed nuclear fragments.
Laser scanning cytometry achieved fluorescent sensitivity comparable to flow cytometry while adding single-cell visual morphological validation on slides.
Chromatin condensation was detectable via increased maximal pixel fluorescence, while early stages of apoptosis were resolved using mitochondrial dyes (rhodamine 123, DiOC6(3), JC-1) and annexin V membrane assays.
Visual examination of slide-bound cells successfully eliminated false positives by distinguishing genuine apoptotic cells from monocytes and macrophages that ingested apoptotic bodies.

Cite This Study

Bedner et al. (1999) studied this question.

synapsesocial.com/papers/6a6c9636a528af2d65c2869e https://doi.org/10.1002/(sici)1097-0320(19990301)35:3<181::aid-cyto1>3.0.co;2-5

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Implication

Methodological review demonstrates multiparametric apoptosis detection in cell models, highlighting how laser scanning cytometry couples fluorescence measurements with visual morphology.

Analysis of apoptosis by laser scanning cytometry

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Cite This Study

Analysis of apoptosis by laser scanning cytometry

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Cite This Study

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