SEM AFM Core in the Houston Methodist Hospital Research Institute 1 article published in JoVE Developmental Biology An Optimized O9-1/Hydrogel System for Studying Mechanical Signals in Neural Crest Cells Tram P. Le*1, Xiaolei Zhao*1, Shannon Erhardt1,2, Jianhua Gu3, Huie Wang3, Tina O. Findley1, Jun Wang1,2 1Department of Pediatrics, McGovern Medical School, The University of Texas Health Science Center at Houston, 2The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences, The University of Texas Health Science Center at Houston, 3SEM AFM Core in the Houston Methodist Hospital Research Institute Detailed step-by-step protocols are described here for studying mechanical signals in vitro using multipotent O9-1 neural crest cells and polyacrylamide hydrogels of varying stiffness.