New method for spectrofluorometer monochromator wavelength calibration

A method is presented for wavelength calibration of spectrofluorometer monochromators. It is based on the distortion that the characteristic absorption bands of glass filters (holmium or didymium oxide), commonly used for calibration of spectrophotometers, introduce in the emitted fluorescence of fluorophores like indole, diphenyl hexatriene, xylene or rhodamine 6G. Those filters or a well characterized absorber with sharp bands like benzene vapor can be used for the same purpose. The wavelength calibration accuracy obtained with this method is better than 0.1 nm, and requires no modification in the geometry of the spectrofluorometer sample compartment.