Most commonly used biological buffers (see Buffer) don’t show a significant fluorescence signal at the red emission wavelengths, relevant for Dianthus experiments. Some substances however can, to test for buffer autofluorescence, load buffer alone into an empty well and measure the well in DI.Control software using the “Quick Start Single-dose template”, and setting the Excitation Power manually to 100 %. Buffer autofluorescence is considered problematic if it is 50% higher than the fluorescence of the labeled target molecule.
To address buffer autofluorescence, change to a non-fluorescent buffer, or remove the fluorescent buffer component. Increasing the target concentration may help but is generally not the recommended solution for this issue.