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Three-Photon Fluorescence Microscopy

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Three-Photon Fluorescence Microscopy

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Three-photon fluorescence (3PF) microscopy can provide a significant advantage over 2PF microscopy in strongly scattering samples such as the mouse brain. The fluorescence from three-photon excitation falls off as 1/z4, where z is the distance away from the focal plane, while the fluorescence from two-photon excitation falls off as 1/z2. Thus, three-photon excitation reduces the background from regions away from the focal plane and improves the signal-to-background ratio by orders of magnitude. This reduction in background is illustrated in Figure 1 where 2PF and 3PF are compared.

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