The dual band fluorescence filter sets contain dual bandpass excitation and emission (barrier) filters, which are incorporated into a single element capable of selectively isolating fluorescence emission from two fluorophores simultaneously. Each of the filter sets is designed for optimal performance with a specific fluorochrome pair, although they are equally effective with alternate pairs of fluorescent probes that have similar absorption and emission spectral profiles. Utilizing precise wavelength band selection, with steep bandpass transitions between reflection and transmission regions, the multiple excitation and emission signals are separated with minimal crossover interference.