1. Using Caenorhabditis elegans as a model for mechanistic insights of craniofacial development
Michael Gruss and Ann K. Corsi
2. Wholemount In-situ Hybridisation (WISH) in Zebrafish Embryos to Analyse Craniofacial DevelopmentNishanthi Mathiyalagan and Sebastian Dworkin
3. Elucidation of Gene Expression Patterns in The Craniofacial Tissues Of Mouse Embryos By Whole-Mount In Situ Hybridization
Joshua B Studdert, Heidi Bildsoe, V Pragathi Masamsetti, and Patrick P L Tam
4. Visualization of the Cartilage and Bone Elements in The Craniofacial Structures by Alcian blue and Alizarin Red Staining
Joshua B Studdert, Heidi Bildsoe, V Pragathi Masamsetti, and Patrick P L Tam
5. Methodology for Free-Floating Organ Culture of Mid-Gestation Maxillary Primordial Tissue
M. E. de Vries and S. Dworkin
6. Methods of Palate Culture in Later Palatogenesis: Elevation, Horizontal Outgrowth, and Fusion
Jack D. Morgan and Jeremy B. A. Green
7. Single cell RNA-seq: Cell Isolation and Data Analysis
Val Yianni and Paul T Sharpe
8. Generating Zebrafish RNA-less Mutant Alleles by Deleting Gene Promoters with CRISPR/Cas9
Priyanka Kumari, Morgan Sturgeon, Gregory Bonde, and Robert A. Cornell
9. Craniofacial Phenomics: Three-Dimensional Assessment of the Size and Shape of Cranial and Dentofacial Structures
Mustafa Mian, Jenny Tan, Robin Yong, Ruth Williams, Agatha Labrinidis, Peter J. Anderson, and Sarbin Ranjitkar
10. Micro-CT-based Bone Microarchitecture Analysis of the Murine Skull
Jenny Tan, Agatha Labrinidis, Ruth Williams, Mustafa Mian, Peter J. Anderson, and Sarbin Ranjitkar
11. Characterization of mammalian in vivo enhancers using mouse transgenesis and CRISPR genome editing
Marco Osterwalder, Stella Tran, Riana D. Hunter, Eman M. Meky, Kianna von Maydell, Anne N. Harrington, Janeth Godoy, Catherine S. Novak, Ingrid Plajzer-Frick, Yiwen Zhu, Jennifer A. Akiyama, Veena Afzal, Evgeny Z. Kvon, Len A. Pennacchio, Diane E. Dickel, and Axel Visel
12. Histological Techniques for Sectioning Bones of the Vertebrate Craniofacial Skeleton
Daniela M. Roth, Lakshmi Puttagunta, and Daniel Graf
13. Engineering Epithelial-Mesenchymal Microtissues to Study Cell-Cell Interactions in Development
Jacob I. Reynolds, Ross A. Vitek, Peter G. Geiger, and Brian P. Johnson
14. Ex Vivo Culture of Human Cranial Suture Cells
Peter J. Anderson
15. Scaffolds for Use in Craniofacial Bone Regeneration
Katherine R. Hixon, Christopher T. Eberlin, Meghana Pendyala, Angela Alarcon de la Lastra, and Scott A. Sell
16. Mandible Explant Assay for The Analysis of Meckel’s Cartilage Development
Sophie Wiszniak and Quenten Schwarz
17. High Resolution Histology for Craniofacial Studies On Zebrafish and Other Teleost Models
A. Huysseune, M. Soenens, J.-Y. Sire, and P.E. Witten
18. Live Imaging the Dynamics Of Mammalian Neural Crest Cell Migration
Emma L. Moore and Paul A. Trainor
19. Salivary Gland Development In Culture Marcia Gaete, Tathyane H. N. Teshima, Lemonia Chatzeli, and Abigail S. Tucker
20. Antenatal Ultrasound Imaging for Analysis of Human Craniosynostosis
Sarah Constantine and Piotr Niznik
This volume explores scientific methodologies currently employed to integrate observational developmental biology, tissue explant and cell-based approaches and genetic/molecular technologies to develop a holistic understanding of craniofacial development. Chapters guide readers through the use of disparate models to study formation of the head and face (c. elegans, zebrafish, mouse, alongside human imaging approaches), together with cell culture, tissue explant and in vivo cell imaging and analysis techniques. At the molecular level, chapters include analysing gene expression using in-situ hybridisation and single-cell RNA-Sequencing (scRNA-SEQ), as well as genetic modification techniques such as CRISPR/Cas9-mediated deletion. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.
Authoritative and cutting-edge, Craniofacial Development: Methods and Protocols aims to be a guide in the field of craniofacial development for senior and new researchers looking to expand their existing research programs to encompass novel techniques.