Prism-based excitation wavelength selection for multicolor fluorescence coincidence measurements

We have designed and constructed a prism setup for multiple-color confocal fluorescence experiments. The prism setup permits easy selection of any color or any combination of colors from a multicolor light source, such as a mixed-gas argon-krypton-ion laser. The selected colors emerging from the prism setup are, by design, optimally overlapped in the focus of a high-numerical-aperture objective, such as that commonly used in single-molecule fluorescence experiments. The various excitation powers can be easily adjusted in this setup. We will exemplify the potential of this setup in single-molecule fluorescence emission spectroscopy.