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|Title:||Whole-mount three-dimensional imaging of internally localized immunostained cells within mouse embryos||Authors:||Yokomizo, T.
|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. https://doi.org/10.1038/nprot.2011.441||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||URI:||http://scholarbank.nus.edu.sg/handle/10635/110798||ISSN:||17542189||DOI:||10.1038/nprot.2011.441|
|Appears in Collections:||Staff Publications|
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