Intravital microscopy: a novel tool to study cell biology in living animals
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Intravital microscopy encompasses various optical microscopy techniques aimed at visualizing biological processes in live animals. In the last decade, the development of non-linear optical microscopy resulted in an enormous increase of in vivo studies, which have addressed key biological questions in fields such as neurobiology, immunology and tumor biology. Recently, few studies have shown that subcellular processes can be imaged dynamically in the live animal at a resolution comparable to that achieved in cell cultures, providing new opportunities to study cell biology under physiological conditions. The overall aim of this review is to give the reader a general idea of the potential applications of intravital microscopy with a particular emphasis on subcellular imaging. An overview of some of the most exciting studies in this field will be presented using resolution as a main organizing criterion. Indeed, first we will focus on those studies in which organs were imaged at the tissue level, then on those focusing on single cells imaging, and finally on those imaging subcellular organelles and structures.
KeywordsIntravital microscopy Non-linear microscopy Live animal imaging Membrane traffic Two-photon microscopy
This research was supported by the Intramural Research Program of the NIH, National Institute of Dental and Craniofacial Research. We apologize to those whose work could not be cited due to space limitations. We would like to thank Dr. Silvio Gutkind, Dr. Julie Donaldson and Dr. Omayma Al-Awar for critical reading of the manuscript and all the members of the Oral and Pharyngeal Cancer Branch for invaluable assistance.
Supplementary movie 3: Volume rendering of the vasculature (500 kDa FITC dextran, green) and the salivary ducts in a live rat (70 kDA Texas red dextran injected into the Wharton’s duct). Excitation wavelength 920 nm (MOV 9232 kb)
Supplementary movie 4: Endocytosis of fluorescently labeled dextran in the salivary glands of live rats. The nuclei are labeled with Hoechst (blue), the vasculature with a 500 kDa FITC dextran (green) and the endosomes with 70 kDa Texas red dextran (red). Time is expressed in min:sec. Excitation wavelength 820 nm (MOV 7925 kb)
Supplementary movie 5: Fusion of lysosomes and dynamics of mitochondria. Lysosomes are labeled with Alexa 488-dextran (green) and mitochondria with the vital dye mitotracker (red). Time lapse performed in single confocal microscopy. (MOV 16306 kb)
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