Improved procedures for immunoferritin labeling of ultrathin frozen sections
Improved procedures for immunoferritin labeling of ultrathin frozen sections
复制标题
超薄冰冻切片免疫铁蛋白标记的改进程序
作者：
K. Tokuyasu;S. Singer
在采用固定的冷冻切片作为细胞内抗原免疫铁蛋白标记的底物时，我们发现常规的戊二醛固定有时只允许对切片进行很少的特异性染色，这要么是因为它使某些蛋白质抗原失活，要么是因为它使抗体染色剂难以接近它们。我们已经开发了几种化学性质较温和的固定程序，可以使样本均匀但不太广泛地交联。在处理更易碎的冷冻切片时，采用这些程序和预防措施，几种系统实现了良好的结构保存和特异性免疫铁蛋白标记。
In employing fixed frozen ultrathin sections as substrates for immunoferritin labeling of intracellular antigens, we have found that conventional glutaraldehyde fixation sometimes permits very little specific staining of the sections, either because it inactivates certain protein antigens, or because it renders them inaccessible to the antibody stains. We have developed several fixation procedures that are chemically milder and allow a uniform but less extensive cross- linking of the specimen. With these procedures and precautions in the handling of the more fragile frozen sections, excellent structural preservation and specific immunoferritin labeling has been achieved with several systems.