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Title: Whole-mount three-dimensional imaging of internally localized immunostained cells within mouse embryos
Authors: Yokomizo, T. 
Yamada-Inagawa, T.
Yzaguirre, A.D.
Chen, M.J.
Speck, N.A.
Dzierzak, E.
Issue Date: Mar-2012
Citation: Yokomizo, T., Yamada-Inagawa, T., Yzaguirre, A.D., Chen, M.J., Speck, N.A., Dzierzak, E. (2012-03). Whole-mount three-dimensional imaging of internally localized immunostained cells within mouse embryos. Nature Protocols 7 (3) : 421-423. ScholarBank@NUS Repository.
Abstract: We describe a three-dimensional (3D) confocal imaging technique to characterize and enumerate rare, newly emerging hematopoietic cells located within the vasculature of whole-mount preparations of mouse embryos. However, the methodology is broadly applicable for examining the development and 3D architecture of other tissues. Previously, direct whole-mount imaging has been limited to external tissue layers owing to poor laser penetration of dense, opaque tissue. Our whole-embryo imaging method enables detailed quantitative and qualitative analysis of cells within the dorsal aorta of embryonic day (E) 10.5-11.5 embryos after the removal of only the head and body walls. In this protocol we describe the whole-mount fixation and multimarker staining procedure, the tissue transparency treatment, microscopy and the analysis of resulting images. A typical two-color staining experiment can be performed and analyzed in ∼6 d. © 2012 Nature America, Inc. All rights reserved.
Source Title: Nature Protocols
ISSN: 17542189
DOI: 10.1038/nprot.2011.441
Appears in Collections:Staff Publications

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