Journal
JOURNAL OF CLINICAL INVESTIGATION
Volume 122, Issue 12, Pages 4439-4446Publisher
AMER SOC CLINICAL INVESTIGATION INC
DOI: 10.1172/JCI65100
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Funding
- Deutsche Jose Carreras Leukamie-Stiftung e.V. [DJCLS R 08/07]
- DFG [SFB TR52, Z2, CRU216, TP4, SCHU 1518/2-1, TR36 Z2]
- Interdisziplinares Zentrum fur Klinische Forschung (IZKF) Wurzburg
- Anton und Petra Ehrmann Stiftung (APES)
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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.
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