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Analysis of cell viability using time-dependent increase in fluorescence intensity.

H K Hamalainen-Laanaya¹, M S Orloff

¹Division of Solid Organ Transplantation, Department of Surgery, University of Rochester Medical Center, Rochester, NY 14623, USA. heli_hamalainenlaanaya@urmc.rochester.edu

Analytical Biochemistry

|July 17, 2012

Summary

This study presents a rapid and cost-effective AlamarBlue assay for assessing primary cell viability. The method accurately measures cell metabolic activity and reduction potential, suitable for high-throughput applications.

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Area of Science:

Cell Biology
Biotechnology

Background:

Resazurin (AlamarBlue) dye reduction is a common cell viability assay.
Current methods are impractical for primary cells due to slow metabolic rates.

Purpose of the Study:

To develop a robust and rapid AlamarBlue assay for primary cell viability.
To enable accurate monitoring of cell metabolic activity and reduction potential.

Main Methods:

Utilized AlamarBlue dye with human mononuclear cells.
Monitored fluorescence intensity increase over several hours.
Assayed cell growth rates, cytotoxicity, and sample quality.

Main Results:

Demonstrated a linear increase in cellular reduction capacity with AlamarBlue.

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The assay proved effective for primary cells, overcoming limitations of slower metabolic activity.

Showcased applicability for assessing frozen cells and pooled donor samples.

Conclusions:

The AlamarBlue assay provides a simple, rapid, and cost-effective method for primary cell viability assessment.
This technique is well-suited for high-throughput studies and immunological research.
The assay can be adapted to monitor reduction potential in various living cells.