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Distinguishing modes of cell death using the ImageStream multispectral imaging flow cytometer.
Thaddeus C George1, David A Basiji, Brian E Hall
1Amnis Corporation, Seattle, Washington 98121, USA. t.george@Amnis.com
Summary
ImageStream 100 Multispectral Imaging Cytometer distinguishes live, necrotic, and apoptotic cells. Unique photometric and morphometric features enable differentiation of cell death stages, improving cell analysis.
Area of Science:
- Cell Biology
- Biotechnology
- Immunology
Background:
- Distinguishing between live, necrotic, and apoptotic cells is crucial for understanding cellular processes.
- Conventional flow cytometry has limitations in differentiating complex cell death stages.
Purpose of the Study:
- To demonstrate the ImageStream 100 Multispectral Imaging Cytometer's capability in discriminating cell death.
- To leverage unique photometric and morphometric features for enhanced cell analysis.
Main Methods:
- Cells (live, necrotic, early/late apoptotic) were analyzed using immunofluorescence microscopy, conventional flow cytometry, and imaging flow cytometry.
- Analysis included single populations and heterogeneous cell mixtures.
Main Results:
- Live and early apoptotic cells were identifiable by both conventional and ImageStream methods.
- ImageStream technology successfully distinguished late apoptotic from necrotic cells, which conventional methods could not.
- Multispectral imaging combined photometric and morphometric data for improved discrimination.
Conclusions:
- Multispectral imaging offers distinct cellular features beyond conventional flow cytometry.
- Coupling morphometric and photometric measures enables precise differentiation of cell viability and death stages.