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J Clin Invest. 2012 Dec;122(12):4439-46. doi: 10.1172/JCI65100. Epub 2012 Nov 12.

Mapping immune processes in intact tissues at cellular resolution.

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1
Department of Medicine II, Würzburg University Hospital, Würzburg, Germany.

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.

PMID:
23143304
PMCID:
PMC3533559
DOI:
10.1172/JCI65100
[Indexed for MEDLINE]
Free PMC Article
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