title: Rapid Acquisition of 3D Images Using High-resolution Episcopic Microscopy.
abstract: High-resolution episcopic microscopic (HREM) technology enables rapid acquisition of high-resolution digital volumetric and three-dimensional (3D) morphometric data. Here, we describe the detailed protocol to image the entire mouse embryo. The protocol consists of four major sections: sample preparation, embedding, image acquisition and finally, 3D visualization. The technology requires specimens to be stained with a fluorescent dye, which can be problematic for large or dense specimens. To overcome this limitation, we have improved the existing protocol to enhance tissue penetration of the dye by pretreating the specimen with a solution containing urea and sodium dodecyl sulfate. The protocol uses only routine laboratory equipment and reagents for easy adaptation in standard laboratory settings. We show that the resulting high-resolution 3D images faithfully recapitulate the detailed morphologic features of the internal organs of mouse embryos, thereby permitting morphometric analyses. Together, we present a detailed and improved protocol using standard laboratory equipment to acquire high-resolution 3D images of small and large sized specimens.
authors.affiliation: Department of Pediatric Surgery, The Second Affiliated Hospital & Yuying Children's Hospital, Wenzhou Medical University; Departments of Urology, Boston Children's Hospital; Department of Surgery, Harvard Medical School.
authors.forename: Haochuan
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Departments of Urology, Boston Children's Hospital; Department of Surgery, Harvard Medical School; Department of Cardiovascular Surgery, Guangdong Cardiovascular Institute, Guangdong General Hospital, Guangdong Academy of Medical Sciences.
authors.forename: JunGang
authors.initials: J
authors.lastname: Huang
authors.affiliation: Departments of Urology, Boston Children's Hospital; Department of Surgery, Harvard Medical School.
authors.forename: Xin
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Cardiovascular Surgery, Guangdong Cardiovascular Institute, Guangdong General Hospital, Guangdong Academy of Medical Sciences.
authors.forename: Ping
authors.initials: P
authors.lastname: Zhu
authors.affiliation: Department of Pediatric Surgery, The Second Affiliated Hospital & Yuying Children's Hospital, Wenzhou Medical University; wmclzr@163.com.
authors.forename: Zhongrong
authors.initials: Z
authors.lastname: Li
authors.affiliation: Departments of Urology, Boston Children's Hospital; Department of Surgery, Harvard Medical School; Sean.Li@childrens.harvard.edu.
authors.forename: Xue
authors.initials: X
authors.lastname: Li
mesh_terms: D000818:Animals; D004622:Embryo, Mammalian; D021621:Imaging, Three-Dimensional; D051379:Mice; D008853:Microscopy / Q000379:methods*; D013048:Specimen Handling