PrestoBlue™ HS Cell Viability Reagent

PrestoBlue HS Cell Viability Reagent is a ready-to-use resazurin-based solution that functions as a cell health indicator by using the reducing power of living cells to quantitatively measure viability. Upon entering living cells, resazurin is reduced to resorufin, a compound that is red in color and highly fluorescent. High background fluorescence caused by various contaminants reduces the performance of resazurin-based reagents. To improve the performance an innovative purification process was developed that removes these contaminants. This highly purified, non-toxic resazurin was used in the standard PrestoBlue formulation, creating the PrestoBlue HS Cell Viability Reagent. Changes in viability can be easily detected using either an absorbance- or fluorescence-based plate reader. The PrestoBlue HS Cell Viability Reagent has been optimized for the fast and reliable detection of mammalian cell viability.

PrestoBlue HS Cell Viability Reagent features include:
• Removal of contaminants from resazurin—displays a >50% reduction in background fluorescence
• Signal-to-background ratio increased by >100%—results in large assay signal window
• Highly sensitive reagent with a linear response—detects as few as 10 cells per well
• Convenient add-and-read format—no mixing, no washing, no cell lysis, and compatible with either fluorescence- or absorbance-based instrumentation
• Optimized for reliable detection of mammalian cell viability in only 10 minutes

Measuring changes in cell viability is a fundamental method for assessing cell health, determining genotoxicity, and evaluating anti-cancer drugs. Cell health can be monitored by detecting changes in several key indicators, including changes to plasma membrane integrity, DNA synthesis, DNA content, enzyme activity, presence of ATP, and cellular reducing environment.

Monitoring changes to the cellular reducing environment or metabolic activity by using resazurin-based reagents is a well-established and reliable indicator of cell viability or death. Upon entering living cells, the cellular reducing environment reduces resazurin to resorufin, a compound that is red in color and highly fluorescent. Viable cells continuously convert resazurin to resorufin, increasing the overall fluorescence of the medium surrounding the cells. Additionally, the conversion of resazurin to resorufin results in a pronounced color change, thus cell viability can also be detected using absorbance-based plate readers.

As a consequence of the synthesis and manufacturing processes, all resazurin-based reagents contain a detectable amount of resorufin contamination. The amount of contamination can vary greatly between sources of the material and manufacturing conditions, contributing to differences in detectable background fluorescence. An innovative process was developed that removes the contaminating resorufin, resulting in the highly pure resazurin used in PrestoBlue HS Cell Viability Reagent. This highly pure resazurin produces a >50% decrease in background fluorescence and a >100% increase in the signal-to-background ratio. Mammalian cell viability can be detected after a 10-minute incubation with PrestoBlue HS Cell Viability Reagent. Since no lysis is required, the diluted PrestoBlue HS solution can be removed and replaced with complete growth medium, for further culturing of the cells.