Three-dimensional microCT imaging of mouse development from early post-implantation to early postnatal stages

In this work, we report the use of iodine-contrast microCT to perform high-throughput 3D morphological analysis of mouse embryos and neonates between embryonic day 8.5 to postnatal day 3, with high spatial resolution up to 3 μm/voxel. We show that mouse embryos at early stages can be imaged either within extra embryonic tissues such as the yolk sac or the decidua without physically disturbing the embryos. This method enables a full, undisturbed analysis of embryo turning, allantois development, vitelline vessels remodeling, yolk sac and early placenta development, which provides increased insights into early embryonic lethality in mutant lines. Moreover, these methods are inexpensive, simple to learn and do not require substantial processing time, making them ideal for high throughput analysis of mouse mutants with embryonic and early postnatal lethality.

[1]  Sonnet J. H. Arlander,et al.  Dial 9-1-1 for DNA damage: the Rad9-Hus1-Rad1 (9-1-1) clamp complex. , 2004, DNA repair.

[2]  A. Joyner,et al.  Deletion of Mouse Rad9 Causes Abnormal Cellular Responses to DNA Damage, Genomic Instability, and Embryonic Lethality , 2004, Molecular and Cellular Biology.

[3]  B. Metscher MicroCT for developmental biology: A versatile tool for high‐contrast 3D imaging at histological resolutions , 2009, Developmental dynamics : an official publication of the American Association of Anatomists.

[4]  Steve D. M. Brown,et al.  High-throughput discovery of novel developmental phenotypes , 2017 .

[5]  Diego Rasskin-Gutman,et al.  High-resolution episcopic microscopy: a rapid technique for high detailed 3D analysis of gene activity in the context of tissue architecture and morphology , 2006, Anatomy and Embryology.

[6]  R. Mark Henkelman,et al.  Design and Implementation of a Custom Built Optical Projection Tomography System , 2013, PloS one.

[7]  R. Mark Henkelman,et al.  4D atlas of the mouse embryo for precise morphological staging , 2015, Development.

[8]  R. Mark Henkelman,et al.  Structural Stabilization of Tissue for Embryo Phenotyping Using Micro-CT with Iodine Staining , 2013, PloS one.

[9]  Richard Baldock,et al.  Bloomsbury report on mouse embryo phenotyping: recommendations from the IMPC workshop on embryonic lethal screening , 2013, Disease Models & Mechanisms.

[10]  James M. Brown,et al.  A bioimage informatics platform for high-throughput embryo phenotyping , 2016, Briefings Bioinform..

[11]  W. Weninger,et al.  Episcopic three-dimensional imaging of embryos. , 2012, Cold Spring Harbor protocols.

[12]  J. Hecksher-Sørensen,et al.  Optical Projection Tomography as a Tool for 3D Microscopy and Gene Expression Studies , 2002, Science.

[13]  J. Epstein,et al.  Rapid 3D Phenotyping of Cardiovascular Development in Mouse Embryos by Micro-CT With Iodine Staining , 2010, Circulation. Cardiovascular imaging.

[14]  B. Metscher MicroCT for comparative morphology: simple staining methods allow high-contrast 3D imaging of diverse non-mineralized animal tissues , 2009, BMC Physiology.

[15]  R. Mark Henkelman,et al.  A novel 3D mouse embryo atlas based on micro-CT , 2012, Development.

[16]  L. Feldkamp,et al.  Practical cone-beam algorithm , 1984 .