Kraehenbuhl J P, Racine L, Griffiths G W
Histochem J. 1980 May;12(3):317-32. doi: 10.1007/BF01006953.
The ability to localize intracellular macromolecules in situ by high resolution techniques has been made possible by the development of antibody labelling of thin sections obtained either from tissues embedded in an hydrophilic matrix, or by ultracryotomy or from conventional plastic embedded tissue. When particle-tagged immunological reagents are used to visualize intracellular antigens, quantitative information can be obtained by combining particle counts with morphometric estimations of compartment volume. Various detection systems have been used successfully for quantitation, which include ferritin-conjugated antibodies, biotin-avidin-ferritin complexes and, more recently, gold-protein A conjugates. Examples of the use of these techniques the localization of secretory proteins in pancreatic exocrine cells, opsin and a large membrane protein in photoreceptor cells of frog retina, and contractile proteins in skeletal muscle are given. Quantitative data obtained by morphometric analysis, both in bovine and rat pancreatic exocrine cells, are compared with values assessed by biochemical methods.
通过对从亲水性基质包埋的组织、超薄切片术或常规塑料包埋组织获得的薄切片进行抗体标记,高分辨率技术已使得在原位定位细胞内大分子成为可能。当使用颗粒标记的免疫试剂来可视化细胞内抗原时,通过将颗粒计数与区室体积的形态测量估计相结合,可以获得定量信息。各种检测系统已成功用于定量,其中包括铁蛋白偶联抗体、生物素-抗生物素蛋白-铁蛋白复合物,以及最近的金-蛋白A偶联物。给出了这些技术在胰腺外分泌细胞中分泌蛋白的定位、青蛙视网膜光感受器细胞中的视蛋白和一种大的膜蛋白,以及骨骼肌中收缩蛋白的应用实例。通过形态测量分析在牛和大鼠胰腺外分泌细胞中获得的定量数据,与通过生化方法评估的值进行了比较。