Becker Klaus, Jährling Nina, Saghafi Saiedeh, Dodt Hans-Ulrich
Cold Spring Harb Protoc. 2013 Jul 1;2013(7):683-4. doi: 10.1101/pdb.prot075820.
This protocol describes the preparation of whole mouse brains and dissected hippocampi for ultramicroscopy (UM), a powerful imaging technique that achieves precise and accurate three-dimensional (3D) reconstructions of intact macroscopic specimens with micrometer resolution. In UM, a specimen in the size range of ∼1-15 mm is illuminated perpendicular to the observation pathway by two thin counterpropagating sheets of laser light. Thus, specimens for UM need to be sufficiently transparent, which requires chemical clearing in most cases. In this protocol, mouse brains and hippocampi are carefully dissected and dehydrated, and then cleared in a solution of benzyl benzoate and benzyl alcohol.
本方案描述了用于超微显微镜（UM）的全小鼠脑和解剖海马体的制备方法，超微显微镜是一种强大的成像技术，能够以微米分辨率对完整的宏观标本进行精确和准确的三维（3D）重建。在超微显微镜中，尺寸范围约为1-15毫米的标本通过两束反向传播的薄激光片垂直于观察路径进行照射。因此，用于超微显微镜的标本需要足够透明，在大多数情况下这需要化学透明处理。在本方案中，小鼠脑和海马体被仔细解剖并脱水，然后在苯甲酸苄酯和苯甲醇的溶液中进行透明处理。
