共焦
共焦显微镜
胚胎
生物
解剖
染色
病理
细胞生物学
生物医学工程
医学
光学
物理
作者
Tomomasa Yokomizo,Tomoko Yamada-Inagawa,Amanda D. Yzaguirre,Michael Chen,Nancy A. Speck,Elaine Dzierzak
出处
期刊:Nature Protocols
[Springer Nature]
日期:2012-02-09
卷期号:7 (3): 421-431
被引量:137
标识
DOI:10.1038/nprot.2011.441
摘要
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.
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