Optimization of fluorescence collection in multiphoton microscopy

Efficient fluorescence collection is critically important to maximize image quality and depth in multiphoton microscopy. Here we present an optimized, large aperture fluorescence collection system for use with the Olympus 20 x 0.95NA objective and two Hamamatsu H7422P-40 GaAsP photomultiplier tubes. Using Zemax optical design software to model the fluorescence intensity distribution and collection geometry, we have designed and constructed an optimized, large aperture detector housing assembly, which provides a significant increase in collected fluorescent signal, image quality, and maximum imaging depth.