Gen5 version required: 1.0 or higher
Basis for the Assay:
The internal environment of proliferating cells is more reduced than that of non-proliferating cells. Compounds such as tetrazolium salts and AlamarBlue™, which can be reduced by cellular metabolic intermediates, can be used to monitor cellular proliferation. The reduction of AlamarBlue results in a measurable color shift, as well as the formation of a fluorescent molecule.
The protocol calls for a fluorescence measurement using a 540/35 excitation filter and a 590/20 emission filter.
The experiment data file does not contain any data.
Plate configuration provides for samples to be run in duplicate. In addition, two blank wells are defined on the plate. In regards to data reduction, only blank well subtraction has been programmed.
The report is configured to provide the (1) raw fluorescence, and (2) blanked fluorescence. The monitoring of AlamarBlue reduction by fluorescence is usually expressed as relative fluorescence emission units (RFU) as a function of the time of incubation.