What variables will effect the linearity of a sample?
Fluorescence intensity is typically directly proportional (linear) to concentration. However, when a concentration is too high, light cannot pass through the sample to cause excitation; thus very high concentrations can have very low fluorescence. This happens when the sample is “quenched”. The fluorometer readings will decrease, even though the concentration is still increasing. Diluting a sample 1:1 or some other convenient ratio will check linearity. If it is linear, the reading will decrease in direct proportion to the dilution.