Gerfen Charles R
National Institutes of Mental Health, Bethesda, Maryland, USA.
Curr Protoc Neurosci. 2003 Aug;Chapter 1:Unit 1.1. doi: 10.1002/0471142301.ns0101s23.
This unit covers some of the basic procedures that are common to a wide range of neuroanatomical protocols. Procedures are provided for the preparation of unfixed, fresh brain tissue as well as for perfusion fixation of animals resulting in fixed neural tissue. A variety of methods for sectioning brains are described, including frozen sectioning in a cryostat, frozen sectioning with a microtome, and sectioning with a vibratome. The choice of sectioning method depends on how the brain has been prepared and what histochemical method is to be used. Three post-sectioning procedures are provided: defatting of slide-mounted sections, thionin staining of the sections, and coating of slides with photographic emulsion for autoradiography. Finally, a procedure is described for subbing slides with gelatin, which is necessary in some protocols in order for the sections to adhere to the slides.
本单元涵盖了广泛的神经解剖学实验方案中一些常见的基本操作步骤。提供了未固定的新鲜脑组织制备方法以及对动物进行灌注固定以获得固定神经组织的方法。描述了多种脑切片方法,包括在低温恒温器中进行冷冻切片、用切片机进行冷冻切片以及用振动切片机进行切片。切片方法的选择取决于脑组织的制备方式以及要使用的组织化学方法。提供了三种切片后操作步骤:对载玻片上的切片进行脱脂、对切片进行硫堇染色以及用照相乳胶覆盖载玻片进行放射自显影。最后,描述了一种用明胶包被载玻片的方法,在某些实验方案中,这是使切片附着在载玻片上所必需的。