Using Beers law, the NanoDrop 1000 software calculates the concentration of samples based on absorbance, pathlength and the molar extinction co-efficient. Therefore accurate pathlength calibration is essential to reporting accurate concentrations. Unlike absorbance spectroscopy, measurement of fluorescence is always reported in RFU (relative fluorescent units) and the actual pathlength-dependence for the fluorospectrometer is much less.
NIST SRM reagents (fluorescein and quinine sulfate) routinely used for assessing fluorescence performance are commercially available. More.
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