Zeller R
European Molecular Biology Laboratory, Heidelberg, Germany.
Curr Protoc Mol Biol. 2001 May;Chapter 14:Unit 14.1. doi: 10.1002/0471142727.mb0101s07.
This unit describes selected methods for fixing and sectioning various forms of biological materials ranging from tissues to single cells. Sections prepared according to these protocols can then be used to examine cell and tissue morphology and in studies involving in situ hybridization, immunohistochemistry, and enzyme histochemistry. The basic protocol describes how tissues and embryos can be prepared for sectioning by fixing in paraformaldehyde followed by embedding in wax, while the alternate protocol describes fixation of suspended or cultured cells. Two support protocols cover preserving and fixing organs by perfusion of whole animals with paraformaldehyde and a procedure for sectioning wax blocks of fixed tissue, plus the subsequent mounting of sections onto prepared or "subbed" glass slides.
本单元介绍了用于固定和切片各种生物材料(从组织到单细胞)的选定方法。按照这些方案制备的切片随后可用于检查细胞和组织形态,以及用于原位杂交、免疫组织化学和酶组织化学等研究。基本方案描述了如何通过在多聚甲醛中固定,然后包埋在石蜡中来制备组织和胚胎用于切片,而替代方案描述了悬浮或培养细胞的固定。两个支持方案涵盖了通过用多聚甲醛灌注全动物来保存和固定器官,以及固定组织蜡块的切片程序,以及随后将切片安装到制备好的或“涂有底膜”的载玻片上。