| | Nikon MicroscopyU: Fluorescence Microscopy - Multiphoton Excitation |
 | | However, because the photophysics governing two-photon excitation is different from that of conventional fluorescence excitation, deleterious effects are occasionally observed with two-photon excitation of certain fluorophores, which in turn limits the applicability of this method for optical sectioning in thin specimens. |
 | | The two-photon microscope allows a greater proportion of the excitation illumination to reach the focal plane because the conditions for absorption are not met outside of the area of light concentration, and out-of-focus absorption is eliminated. |
 | | For the one-photon excitation mechanism (confocal microscopy, Figure 5(a)), the intensity exhibits a steady decrease with penetration depth as the excitation light is increasingly absorbed as it reaches the deeper focal planes. |
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