Tokuyasu K T, Singer S J
J Cell Biol. 1976 Dec;71(3):894-906. doi: 10.1083/jcb.71.3.894.
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.
在使用固定的冷冻超薄切片作为细胞内抗原免疫铁蛋白标记的底物时,我们发现传统的戊二醛固定有时会使切片的特异性染色很少,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无法接近这些抗原。我们已经开发了几种化学性质较温和的固定方法,这些方法能使标本进行均匀但程度较低的交联。通过这些方法以及在处理更脆弱的冷冻切片时采取的预防措施,使用几种系统都实现了出色的结构保存和特异性免疫铁蛋白标记。