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Phosphotungstic acid‐enhanced microCT: Optimized protocols for embryonic and early postnatal mice
Author(s) -
Lesciotto Kate M.,
Motch Perrine Susan M.,
Kawasaki Mizuho,
Stecko Timothy,
Ryan Timothy M.,
Kawasaki Kazuhiko,
Richtsmeier Joan T.
Publication year - 2020
Publication title -
developmental dynamics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.634
H-Index - 141
eISSN - 1097-0177
pISSN - 1058-8388
DOI - 10.1002/dvdy.136
Subject(s) - biology , embryonic stem cell , staining , soft tissue , phosphotungstic acid , in situ , visualization , anatomy , biomedical engineering , microbiology and biotechnology , pathology , computer science , biochemistry , chemistry , data mining , medicine , gene , catalysis , genetics , organic chemistry
Background Given the need for descriptive and increasingly mechanistic morphological analyses, contrast‐enhanced microcomputed tomography (microCT) represents perhaps the best method for visualizing 3D biological soft tissues in situ. Although staining protocols using phosphotungstic acid (PTA) have been published with beautiful visualizations of soft tissue structures, these protocols are often aimed at highly specific research questions and are applicable to a limited set of model organisms, specimen ages, or tissue types. We provide detailed protocols for micro‐level visualization of soft tissue structures in mice at several embryonic and early postnatal ages using PTA‐enhanced microCT. Results Our protocols produce microCT scans that enable visualization and quantitative analyses of whole organisms, individual tissues, and organ systems while preserving 3D morphology and relationships with surrounding structures, with minimal soft tissue shrinkage. Of particular note, both internal and external features of the murine heart, lungs, and liver, as well as embryonic cartilage, are captured at high resolution. Conclusion These protocols have broad applicability to mouse models for a variety of diseases and conditions. Minor experimentation in the staining duration can expand this protocol to additional age groups, permitting ontogenetic studies of internal organs and soft tissue structures within their 3D in situ position.