title: Intravital two-photon imaging of the gastrointestinal tract.
abstract: Live imaging of the gastrointestinal tract with two-photon microscopy (TPM) has proven to be a useful tool for mucosal immunologists. It provides deep penetration of live tissues with reduced phototoxicity and photobleaching and thus excels in deciphering dynamic immunological processes that require cell motility and last minutes through hours. The few studies that employed this technique in the gut have uncovered new aspects of mucosal immunity. They focused mainly on adaptive immunity in the small intestine and exposed the details of important interactions among several epithelial and hematopoietic cell types. TPM can be employed either on explanted tissue or intravitally, as has been practiced in our lab. Intravital TPM preserves physiological conditions more faithfully, but it is a demanding technique that requires dedicated personnel. To achieve success, the peristaltic motility of the intestine must be curbed, surgical and photonic damage must be minimized, and tissue degradation must be delayed and controlled for. Here we briefly review published studies that employed intravital TPM in the gut, describe our own technique for imaging the intestinal Peyer's patches (PPs) and villi, and present some observations we made using this technique.
authors: Department of Immunology, The Weizmann Institute of Science, Rehovot, 76100, Israel.
authors: Masha
authors: M
authors: Kolesnikov
authors: Department of Microbiology and Immunology, Harvard Medical School, Boston, MA 02115, USA.
authors: Julia
authors: J
authors: Farache
authors: Department of Immunology, The Weizmann Institute of Science, Rehovot, 76100, Israel. Electronic address: shakhar@weizmann.ac.il.
authors: Guy
authors: G
authors: Shakhar
mesh_terms: D019264:Adoptive Transfer; D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology; D005456:Fluorescent Dyes; D018928:Immunity, Mucosal; D007413:Intestinal Mucosa / Q000098:blood supply / Q000166:cytology* / Q000276:immunology; D007421:Intestine, Small / Q000098:blood supply / Q000166:cytology* / Q000276:immunology; D007694:Killer Cells, Natural / Q000166:cytology; D008264:Macrophages / Q000166:cytology; D051379:Mice; D008810:Mice, Inbred C57BL; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D010581:Peyer's Patches / Q000098:blood supply / Q000166:cytology* / Q000276:immunology; D013601:T-Lymphocytes / Q000166:cytology
chemical_list: D005456:Fluorescent Dyes
keywords: In vivo imaging; Lamina propria; Mucosal immunology; Peyer's patch; Small intestine; Two-photon microscopy