Hi Judit,

I would not say it is confusing, rather I would say are we interpreting the variables used?

The Alamar Blue, alongside other cell viability assays, directly measures the reducing potential of metabolic active cells, mainly through the reductase (e.g mitochrondrial or cytoplasmatic) activities. Ascorbic acid is a huge antioxidant and reducing agent. You could check if this interaction is somewhat interfering with your results, through a blank sample. For example, you take a sample-set of wells and add Alamar blue reagent and your used dose of Ascorbic acid, to check if there is fluorescence there (probably it will, since ascorbic acid will reduce the oxidized reagent).

There is no way to look "only at cell viability", since the viability in this assay is directly correlated to the reducing potential. But taking the blank values off your treatments, maybe you find something.

Hope it helps,

Regards.

Thank you very much, we will try it!