High-Speed Imaging Using Multiphoton Excitation Microscopy

Abstract

Abstract Multiphotonmicroscopy (Denk et al., 1990; Helmchen & Denk, 2002; So et al., 2000; Zipfel et al., 2003) produces three-dimensionally resolved images based on nonlinear optical interactions localized at the focus of a microscope objective. Multiphoton microscopy in the fluorescence mode is now the method of choice for in vivo deep tissue microscopic imaging because of its subcellular resolution, minimal phototoxicity, and excellent tissue penetration depth. For many tissues, penetration depths from a few hundred microns up to a millimeter have been reported (Helmchen & Denk, 2005). Multiphoton microscopy has become an invaluable tool in biomedical studies such as neuronal plasticity (Grutzendler et al., 2002; Lee et al., 2005; Lendvai et al., 2000), angiogenesis in solid tumors (Padera et al., 2002), and transdermal drug delivery (Yu et al., 2001).