Brorson S H, Skjørten F
Department of Pathology, Ullevål Hospital, Oslo, Norway.
Micron. 1996 Jun-Aug;27(3-4):203-9. doi: 10.1016/0968-4328(96)00033-9.
The purpose of this study was to predict the ratio of immunogold labeling of deplasticized epoxy sections and LR-White sections on the basis of theoretical considerations. Tissues used in experiments were pancreas, pituitary, kidney, thyroid, and fibrin. Antigens used as test proteins were glucagon, somatostatin, thyroglobulin, chromogranin A, ACTH (= Adrenocorticotropt hormone), amyloid A, and fibrinogen. These are proteins of different sizes. The quotient labelingdeplasticized/labelingLR-white was deduced theoretically and compared to measurements based on immunogold experiments to obtain a theoretical model with acceptable correlation to the measurements. This study describes a theory--expressed mathematically--for what happens at the molecular level in immunoelectron microscopy at the surface of deplasticized epoxy sections and acrylic sections. The theory explains why we normally get about the same amount of immunogold labeling using LR-White sections (acrylic resin) and deplasticized epoxy sections. Taking the nuances into account, the theory indicates increased usefulness of deplasticized epoxy sections when the diameter of the protein carrying the epitopes decreases.
本研究的目的是基于理论考量预测去塑料化环氧树脂切片和LR-White切片的免疫金标记比率。实验中使用的组织有胰腺、垂体、肾脏、甲状腺和纤维蛋白。用作测试蛋白的抗原有胰高血糖素、生长抑素、甲状腺球蛋白、嗜铬粒蛋白A、促肾上腺皮质激素(ACTH)、淀粉样蛋白A和纤维蛋白原。这些是不同大小的蛋白质。从理论上推导出去塑料化标记/LR-White标记的商,并与基于免疫金实验的测量结果进行比较,以获得与测量结果具有可接受相关性的理论模型。本研究描述了一种用数学表达的理论,用于解释在去塑料化环氧树脂切片和丙烯酸切片表面的免疫电子显微镜中分子水平上发生的情况。该理论解释了为什么我们通常使用LR-White切片(丙烯酸树脂)和去塑料化环氧树脂切片时会得到大致相同数量的免疫金标记。考虑到细微差别,该理论表明当携带表位的蛋白质直径减小时,去塑料化环氧树脂切片的实用性会增加。