Herken R, Fussek M, Thies M
Zentrum Anatomie, Abteilung Histologie, Göttingen, Federal Republic of Germany.
Histochemistry. 1988;89(3):277-82. doi: 10.1007/BF00493152.
This work describes a technique which permits study of the postembedding lectin histochemistry for WGA-binding sites at the light and electron microscopical level on the same resin embedded tissue without removing or etching of the resin. Unfixed kidney pieces or kidney pieces fixed in 4% formaldehyde were embedded in the hydrophilic polyhydroxy aromatic resins LR-Gold and LR-White, following dehydration in up to 70% ethanol, 90% ethanol or 100% ethanol. LR-Gold was cryopolymerised at -25 degrees C using the light sensitive initiator benzil, whereas LR-White was heat-cured at +50 degrees C. The localisation of WGA-binding sites at the light microscopical level was investigated using FITC-labelled WGA. The ultrastructural localisation of WGA-binding sites was performed using 15 nm gold-labelled WGA. The best fluorescence staining results were obtained on fixed or unfixed tissue dehydrated in up to 70% ethanol and embedded in LR-Gold. At the ultrastructural level, the best staining results for WGA-binding sites were seen on tissue sections, dehydrated in up to 90% ethanol prior to embedding in LR-Gold.
这项工作描述了一种技术,该技术允许在不除去或蚀刻树脂的情况下,在光镜和电镜水平上对同一树脂包埋组织中的WGA结合位点进行包埋后凝集素组织化学研究。未固定的肾组织块或用4%甲醛固定的肾组织块,在依次用高达70%乙醇、90%乙醇或100%乙醇脱水后,包埋于亲水性多羟基芳香树脂LR-金和LR-白中。LR-金使用光敏引发剂二苯乙二酮在-25℃下进行低温聚合,而LR-白在+50℃下进行热固化。使用异硫氰酸荧光素标记的WGA研究光镜水平上WGA结合位点的定位。使用15纳米金标记的WGA进行WGA结合位点的超微结构定位。在高达70%乙醇中脱水并包埋于LR-金中的固定或未固定组织上获得了最佳荧光染色结果。在超微结构水平上,在包埋于LR-金之前在高达90%乙醇中脱水的组织切片上观察到WGA结合位点的最佳染色结果。