Multiphoton microscopy in biological research

From its conception a decade ago, multiphoton microscopy has evolved from a photonic novelty to an indispensable tool for gleaning information from subcellular events within organized tissue environments. Its relatively deep optical penetration has recently been exploited for subcellularly resolved investigations of disease models in living transgenic mice. Its enhanced spectral accessibility enables aberration-free imaging of fluorescent molecules absorbing in deep-UV energy regimes with simultaneous imaging of species having extremely diverse emission spectra. Although excited fluorescence is the primary signal for multiphoton microscopy, harmonic generation by multiphoton scattering processes are also valuable for imaging species with large anharmonic modes, such as collagen structures and membrane potential sensing dyes.