Cornelese-ten Velde I, Prins F A
Department of Cytochemistry and Cytometry, University of Leiden, The Netherlands.
Histochemistry. 1990;94(1):61-71. doi: 10.1007/BF00266791.
One simple post-embedding method for combined light- and electron microscopy is presented. Different types of antigens in normal rat and mouse kidneys as well as in tissues from cases of experimental induced nephritis were stained after Lowicryl K4M embedding by an immunogold (silver) method. The (silver-enhanced) gold particles were visualized by light microscopy, e.g. bright-field (BFM)- and reflection contrast (RCM) microscopy, as well as by electron microscopy. The potentials of RCM visualization in this field were investigated, resulting in the successful detection of colloidal gold (15 nm) particles, or silver enhanced gold particles, on ultrathin sections. Furthermore, an increased detection sensitivity of RCM compared with BFM together with an increase in the sensitivity of the immunostaining by RCM visualization was found. The different ways to use RCM, alone or in combination with bright-field- or phase contrast microscopy for visualization of plastic sections varying in thickness, type of plastic and staining, are discussed.
本文介绍了一种用于光镜和电镜联合观察的简单包埋后方法。采用免疫金（银）法对正常大鼠和小鼠肾脏以及实验性诱导肾炎病例组织中的不同类型抗原进行Lowicryl K4M包埋后染色。通过光学显微镜，如明场（BFM）和反射对比（RCM）显微镜，以及电子显微镜对（银增强的）金颗粒进行观察。研究了RCM在该领域的观察潜力，成功地在超薄切片上检测到了胶体金（15nm）颗粒或银增强金颗粒。此外，发现RCM与BFM相比检测灵敏度提高，并且通过RCM观察可提高免疫染色的灵敏度。讨论了单独使用RCM或与明场或相差显微镜联合使用RCM观察不同厚度、塑料类型和染色的塑料切片的不同方法。
