Determining viable cell numbers in cell culture is an essential measure across various biological fields.
Traditionally, manual counting with trypan blue was the standard. While automated cell counters offer convenience, they might not be ideal for high-throughput applications.
Enzyme-based methods, such as dehydrogenase assays, can provide a more scalable solution.
By measuring enzymatic activity, researchers can indirectly assess cell viability. ATP-based luminescence measurements offer a direct and sensitive approach.
Using thermostable firefly luciferase, this method provides a stable readout for extended periods.
Compared to other techniques, ATP-based luminescence is generally more sensitive, reliable, and easier to implement, making it well-suited for high-throughput screening.

LIVE-Step™ Cell Assay System

The LIVE-Step™ Cell Assay System is designed for high-throughput, homogeneous, sensitive luminescence quantification of metabolically active, viable mammalian cells.
It consists of a single ready-to-use solution to lyse the cells and measure the amount of ATP present in culture in one step.
Detection is linear up to >100,000 suspension cells and >50,000 adherent cells with an R2 value greater than 0.99.
The signal is stable for longer than 5 hours (T1/2 >300 min) allowing for experimental flexibility.
This product is compatible with multiple media formulations and the presence of phenol red, and stable after multiple freeze-thaw cycles.
 
Figure 1: Luciferin activity in the presence of ATP.
 
Luciferin in the presence of Mg2+, O2 and ATP released from lysed metabolically active cells generates a luminescence signal.
The signal generated is proportional to the number of metabolically active viable cells that were present in culture prior to lysis.

 

Figure 2: LIVE-Step™ Cell Assay System protocol overview.
 
The addition of the LIVE-Step™ Cell Solution to wells containing mammalian cells results in cell lysis and produces the luciferase-mediated reaction shown in Figure 1.
The signal generated is then measured in a luminometer. The signal is proportional to the fraction of metabolically active, viable cells present in culture prior to lysis.
 
 
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