The methylation reactions were followed by monitoring the increase in fluorescence using BioTek Synergy2 plate reader with 360/40 nm excitation filter and 528/20 nm emission filter for 20 min in 384 - well format. (thesgc.org)
The practical brightness takes all application specific parameters into account, including the specs of your microscope (excitation wavelength, available emission filters, and detector sensitivity) and the biological system (temperature, prokaryote versus eukaryote, background fluorescence). (blog.addgene.org)
The practical brightness assay (employing co-expressing fluorescent proteins or using endogenous tagged genes) can be used to verify performance under different imaging conditions (e.g. different emission filters) or different set - ups. (blog.addgene.org)