Viability assays for cells in culture

Area of Science:

• Cell Biology
• Biotechnology
• Assay Development

Background:

• Manual cell counting via microscopy is sensitive but time-consuming and costly.
• Computerized viability assays offer speed and objectivity but require equipment investment.
• Existing methods necessitate a comparison of automated approaches for efficiency.

Purpose of the Study:

• To evaluate three computerized cell viability assays: two infrared and one luminescent.
• To compare their performance against manual cell counting and each other.
• To assess their utility in reducing experimental time and improving data reliability.

Main Methods:

• Utilized two infrared assays with an Odyssey Imager: DRAQ5/Sapphire staining and In-Cell Western assays (targeting cytoskeletal proteins).
• Employed a luminescent ATP assay using reduced reagent volume for cost-effectiveness.
• Correlated assay measurements with cell numbers and assessed sensitivity and linearity.

Main Results:

• All three computerized assays demonstrated linear measurements correlating with cell numbers.
• Assays circumvented microscopy, sampled entire wells, and reduced sampling error.
• Measurements were completed within one day, enabling higher experimental throughput.

Conclusions:

• Computerized viability assays are a valuable investment despite initial equipment costs.
• All assays, including manual counts, have inherent limitations and assumptions.
• Combining multiple assays provides a comprehensive understanding of cellular viability and function.