The OptoSplit was designed as a convenient, inexpensive solution to simultaneous dual wavelength imaging. It has the unique feature of a rotating mirror cradle, which allows adjustable spatial separation, to facilitate pixel-to-pixel image registration. The OptoSplit can be used with any image capture software and the processing carried out manually off-line. However device drivers are included in many commercial software packages to assist image registration and to allow real-time or off-line ratioing and fluorescence overlays. The simple and accessible design makes the OptoSplit II an excellent platform for such demanding applications as dual polarization imaging. While optimized for coupling to a scientific microscope, the OptoSplit can also be used with camera lenses or any other system of lenses which produce an image plane of suitable size.