Quantitative assessment of cell viability based on flow cytometry and microscopy

Area of Science:

• Immunology
• Cell Biology
• Biotechnology

Background:

• Accurate cell viability assessment is crucial in biological research and diagnostics.
• Flow cytometry and microscopy are common methods for determining cell viability.
• Standardization and comparison of these methods are essential for reliable results.

Purpose of the Study:

• To directly compare the accuracy and agreement of flow cytometry and microscopy for cell viability determination.
• To evaluate the performance of these methods using specific fluorescence viability stains.
• To assess the consistency of results across different cell types, including monocytes and CD34-positive cells.

Main Methods:

• Peripheral blood monocytes were isolated using density gradients and enriched via magnetic-activated cell sorting (MACS) or fluorescence-activated cell sorting (FACS) targeting CD14.
• Identical cell samples were analyzed using both flow cytometry and microscopy.
• Cell viability was determined using calcein acetoxy-methyl-ester (live stain) and ethidium homodimer-1 (dead stain).
• Over 1,000 cells were analyzed per sample to ensure statistical significance.

Main Results:

• Flow cytometry and microscopy demonstrated good agreement in cell viability measurements.
• Average differences in viability were 2.7 ± 1.4% for live staining and 1.7 ± 1.2% for dead staining.
• These deviations were comparable to the inherent measurement uncertainty, indicating equivalent performance.
• Consistent results were observed for both monocytes and MACS-enriched CD34-positive cells.

Conclusions:

• Flow cytometry and microscopy provide comparable and reliable cell viability data.
• Both methods are suitable for assessing cell viability in immunological and cell sorting applications.
• The choice between flow cytometry and microscopy can be based on experimental needs and available resources without compromising accuracy.