title: Mapping immune processes in intact tissues at cellular resolution.
abstract: Understanding the spatiotemporal changes of cellular and molecular events within an organism is crucial to elucidate the complex immune processes involved in infections, autoimmune disorders, transplantation, and neoplastic transformation and metastasis. Here we introduce a novel multicolor light sheet fluorescence microscopy (LSFM) approach for deciphering immune processes in large tissue specimens on a single-cell level in 3 dimensions. We combined and optimized antibody penetration, tissue clearing, and triple-color illumination to create a method for analyzing intact mouse and human tissues. This approach allowed us to successfully quantify changes in expression patterns of mucosal vascular addressin cell adhesion molecule-1 (MAdCAM-1) and T cell responses in Peyer's patches following stimulation of the immune system. In addition, we employed LSFM to map individual T cell subsets after hematopoietic cell transplantation and detected rare cellular events. Thus, we present a versatile imaging technology that should be highly beneficial in biomedical research.
authors.affiliation: Department of Medicine II, Würzburg University Hospital, Würzburg, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Brede
authors.forename: Mike
authors.initials: M
authors.lastname: Friedrich
authors.forename: Ana-Laura
authors.initials: AL
authors.lastname: Jordán-Garrote
authors.forename: Simone S
authors.initials: SS
authors.lastname: Riedel
authors.forename: Carina A
authors.initials: CA
authors.lastname: Bäuerlein
authors.forename: Katrin G
authors.initials: KG
authors.lastname: Heinze
authors.forename: Tobias
authors.initials: T
authors.lastname: Bopp
authors.forename: Stephan
authors.initials: S
authors.lastname: Schulz
authors.forename: Anja
authors.initials: A
authors.lastname: Mottok
authors.forename: Carolin
authors.initials: C
authors.lastname: Kiesel
authors.forename: Katharina
authors.initials: K
authors.lastname: Mattenheimer
authors.forename: Miriam
authors.initials: M
authors.lastname: Ritz
authors.forename: Viktoria
authors.initials: V
authors.lastname: von Krosigk
authors.forename: Andreas
authors.initials: A
authors.lastname: Rosenwald
authors.forename: Hermann
authors.initials: H
authors.lastname: Einsele
authors.forename: Robert S
authors.initials: RS
authors.lastname: Negrin
authors.forename: Gregory S
authors.initials: GS
authors.lastname: Harms
authors.forename: Andreas
authors.initials: A
authors.lastname: Beilhack
mesh_terms: D056704:Adaptive Immunity; D000818:Animals; D006086:Graft vs Host Disease / Q000473:pathology; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008856:Microscopy, Fluorescence; D059010:Single-Cell Analysis; D051598:Whole Body Imaging