Continuous intact cell detection and viability determination by CE with dual-wavelength detection

Area of Science:

• Analytical Chemistry
• Cell Biology
• Biotechnology

Background:

• Accurate cell viability assessment is crucial in biological research and drug development.
• Conventional methods like manual trypan blue staining are time-consuming and prone to human error.
• There is a need for automated, high-throughput methods for individual cell analysis.

Purpose of the Study:

• To develop and validate a capillary electrophoresis (CE) method for continuous, individual cell viability determination.
• To compare the performance of the CE method with conventional viability assays.
• To demonstrate the utility of the CE method in cytotoxicity studies.

Main Methods:

• Cells were stained with trypan blue and fixed with formaldehyde.
• Continuous cell introduction into the capillary was achieved using electroosmotic flow (EOF).
• Dual-wavelength ultraviolet-visible detection was employed, with absorbance at 590 nm used for viability assessment.

Main Results:

• A clear cut-off absorbance of 2 milli-absorbance units at 590 nm distinguished live from dead Hela cells.
• The CE method showed excellent correlation (R²=0.9623) with conventional trypan blue staining.
• Results from methylmercury cytotoxicity assays were consistent across CE, trypan blue, and MTT assays.

Conclusions:

• The developed CE method provides an automated and accurate approach for continuous cell viability determination.
• This method overcomes limitations of manual counting, reducing bias and improving efficiency.
• The CE technique is suitable for individual cell analysis and cytotoxicity screening.