title: Intravital microscopy: a practical guide on imaging intracellular structures in live animals.
abstract: Intravital microscopy is an extremely powerful tool that enables imaging several biological processes in live animals. Recently, the ability to image subcellular structures in several organs combined with the development of sophisticated genetic tools has made possible extending this approach to investigate several aspects of cell biology. Here we provide a general overview of intravital microscopy with the goal of highlighting its potential and challenges. Specifically, this review is geared toward researchers that are new to intravital microscopy and focuses on practical aspects of carrying out imaging in live animals. Here we share the know-how that comes from first-hand experience, including topics such as choosing the right imaging platform and modality, surgery and stabilization techniques, anesthesia and temperature control. Moreover, we highlight some of the approaches that facilitate subcellular imaging in live animals by providing numerous examples of imaging selected organelles and the actin cytoskeleton in multiple organs.
authors.affiliation: Intracellular Membrane Trafficking Unit; Oral and Pharyngeal Cancer Branch; National Institute of Dental and Craniofacial Research; National Institutes of Health; Bethesda, MD USA; Department of Biology; University of North Carolina at Chapel Hill; Chapel Hill, NC USA.
authors.forename: Andrius
authors.initials: A
authors.lastname: Masedunskas
authors.affiliation: Intracellular Membrane Trafficking Unit; Oral and Pharyngeal Cancer Branch; National Institute of Dental and Craniofacial Research; National Institutes of Health; Bethesda, MD USA; Department of Chemical and Biochemical Engineering; Rutgers University; Piscataway, NJ USA; Department of Biomedical Engineering; Rutgers University; Piscataway, NJ USA.
authors.forename: Oleg
authors.initials: O
authors.lastname: Milberg
authors.affiliation: Intracellular Membrane Trafficking Unit; Oral and Pharyngeal Cancer Branch; National Institute of Dental and Craniofacial Research; National Institutes of Health; Bethesda, MD USA.
authors.forename: Natalie
authors.initials: N
authors.lastname: Porat-Shliom
authors.affiliation: Intracellular Membrane Trafficking Unit; Oral and Pharyngeal Cancer Branch; National Institute of Dental and Craniofacial Research; National Institutes of Health; Bethesda, MD USA.
authors.forename: Monika
authors.initials: M
authors.lastname: Sramkova
authors.affiliation: Intracellular Membrane Trafficking Unit; Oral and Pharyngeal Cancer Branch; National Institute of Dental and Craniofacial Research; National Institutes of Health; Bethesda, MD USA.
authors.forename: Tim
authors.initials: T
authors.lastname: Wigand
authors.affiliation: Intracellular Membrane Trafficking Unit; Oral and Pharyngeal Cancer Branch; National Institute of Dental and Craniofacial Research; National Institutes of Health; Bethesda, MD USA.
authors.forename: Panomwat
authors.initials: P
authors.lastname: Amornphimoltham
authors.affiliation: Intracellular Membrane Trafficking Unit; Oral and Pharyngeal Cancer Branch; National Institute of Dental and Craniofacial Research; National Institutes of Health; Bethesda, MD USA.
authors.forename: Roberto
authors.initials: R
authors.lastname: Weigert
mesh_terms: D008841:Actin Cytoskeleton; D000818:Animals; D008853:Microscopy / Q000379:methods*; D015388:Organelles