Herken R, Fussek M, Barth S, Götz W
Abteilung Histologie, Georg-August Universität Göttingen, Germany.
Histochem J. 1988 Aug;20(8):427-32. doi: 10.1007/BF01002428.
This work describes a method for the immunolocalization of laminin on 1 micron-thick tissue sections using a postembedding immunofluorescence technique. Embedding of unfixed or formaldehyde-fixed mouse renal cortex in either of the acrylic resins LR-White or LR-Gold permitted reliable postembedding immunofluorescence staining for laminin. LR-White was heat-cured at 50 degrees C whereas LR-Gold was polymerized at -25 degrees C. A stronger immunostaining for laminin was obtained from tissue embedded in polymerized LR-Gold compared with the staining from tissue embedded in LR-White. Prerequisites for adequate postembedding immunostaining are the partial dehydration of the tissue (maximum ethanol concentration, 70%) and pepsin treatment of the tissue sections prior to performing the immunostaining reactions.
这项工作描述了一种使用包埋后免疫荧光技术在1微米厚的组织切片上进行层粘连蛋白免疫定位的方法。将未固定或甲醛固定的小鼠肾皮质包埋在丙烯酸树脂LR-White或LR-Gold中,均可对层粘连蛋白进行可靠的包埋后免疫荧光染色。LR-White在50℃加热固化,而LR-Gold在-25℃聚合。与LR-White包埋的组织染色相比,LR-Gold聚合包埋的组织对层粘连蛋白的免疫染色更强。进行充分的包埋后免疫染色的前提条件是组织的部分脱水(最大乙醇浓度为70%)以及在进行免疫染色反应之前对组织切片进行胃蛋白酶处理。