title: In vivo imaging of tracheal epithelial cells in mice during airway regeneration.
abstract: Many human lung diseases, such as asthma, chronic obstructive pulmonary disease, bronchiolitis obliterans, and cystic fibrosis, are characterized by changes in the cellular composition and architecture of the airway epithelium. Intravital fluorescence microscopy has emerged as a powerful approach in mechanistic studies of diseases, but it has been difficult to apply this tool for in vivo respiratory cell biology in animals in a minimally invasive manner. Here, we describe a novel miniature side-view confocal probe capable of visualizing the epithelium in the mouse trachea in vivo at a single-cell resolution. We performed serial real-time endotracheal fluorescence microscopy in live transgenic reporter mice to view the three major cell types of the large airways, namely, basal cells, Clara cells, and ciliated cells. As a proof-of-concept demonstration, we monitored the regeneration of Clara cells over 18 days after a sulfur dioxide injury. Our results show that in vivo tracheal microscopy offers a new approach in the study of altered, regenerating, or metaplastic airways in animal models of lung diseases.
authors.affiliation: Harvard Medical School, Massachusetts General Hospital, Boston, MA, USA.
authors.forename: Jun Ki
authors.initials: JK
authors.lastname: Kim
authors.forename: Vladimir
authors.initials: V
authors.lastname: Vinarsky
authors.forename: John
authors.initials: J
authors.lastname: Wain
authors.forename: Rui
authors.initials: R
authors.lastname: Zhao
authors.forename: Keehoon
authors.initials: K
authors.lastname: Jung
authors.forename: Jinwoo
authors.initials: J
authors.lastname: Choi
authors.forename: Adam
authors.initials: A
authors.lastname: Lam
authors.forename: Ana
authors.initials: A
authors.lastname: Pardo-Saganta
authors.forename: Sylvie
authors.initials: S
authors.lastname: Breton
authors.forename: Jayaraj
authors.initials: J
authors.lastname: Rajagopal
authors.forename: Seok Hyun
authors.initials: SH
authors.lastname: Yun
mesh_terms: D000818:Animals; D058948:Cell Tracking; D004847:Epithelial Cells / Q000473:pathology*; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D055370:Lung Injury / Q000139:chemically induced / Q000473:pathology; D051379:Mice; D008856:Microscopy, Fluorescence / Q000295:instrumentation; D011401:Promoter Regions, Genetic; D012038:Regeneration; D013234:Stem Cells / Q000473:pathology; D013458:Sulfur Dioxide; D059008:Time-Lapse Imaging; D014132:Trachea / Q000473:pathology / Q000503:physiopathology*; D025262:Vacuolar Proton-Translocating ATPases / Q000235:genetics
chemical_list: C485184:enhanced green fluorescent protein; D013458:Sulfur Dioxide; D049452:Green Fluorescent Proteins; C480628:Atp6v1b1 protein, mouse; D025262:Vacuolar Proton-Translocating ATPases