title: Two-photon excitation microscopy for the study of living cells and tissues.
abstract: Two-photon excitation microscopy is an alternative to confocal microscopy that provides advantages for three-dimensional and deep tissue imaging. This unit will describe the basic physical principles behind two-photon excitation and discuss the advantages and limitations of its use in laser-scanning microscopy. The principal advantages of two-photon microscopy are reduced phototoxicity, increased imaging depth, and the ability to initiate highly localized photochemistry in thick samples. Practical considerations for the application of two-photon microscopy will then be discussed, including recent technological advances. This unit will conclude with some recent applications of two-photon microscopy that highlight the key advantages over confocal microscopy and the types of experiments which would benefit most from its application.
authors.affiliation: University of Colorado, Anschutz Medical Campus, Aurora, Colorado.
authors.forename: Richard K P
authors.initials: RKP
authors.lastname: Benninger
authors.affiliation: Vanderbilt University Medical Center, Nashville, Tennessee.
authors.forename: David W
authors.initials: DW
authors.lastname: Piston
mesh_terms: D000818:Animals; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods; D008853:Microscopy / Q000295:instrumentation / Q000379:methods*; D055095:Optics and Photonics / Q000379:methods*; D011506:Proteins / Q000378:metabolism
chemical_list: D011506:Proteins