Pixel shift results when a filter in an imaging path (the emitter and/or dichroic beamsplitter in a fluorescence microscope) deviates the light rays to cause a shift of the image detected on a high-resolution monochrome digital camera. This shift becomes problematic when two or more images of the same object are acquired using different filter sets and then overlaid to simultaneously view fluorescence from multiple fluorophores. Images produced by different fluorophores (and different filter sets) will not be accurately correlated or combined because each image is shifted by a different amount depending on the wedge angles found in each filter set. To eliminate pixel shift, BrightLine ZERO and Avant ZERO filter sets are manufactured and tested to exacting tolerances to ensure accurate image registration when combining multiple images.