Nitsch R, Klauer G
Institute of Anatomy, Johann Wolfgang Goethe University, Frankfurt am Main, Federal Republic of Germany.
Histochemistry. 1989;92(6):459-65. doi: 10.1007/BF00524757.
Perfusion-fixed tissue blocks were incubated in high molar sucrose solutions, shock frozen in melting isopentane, and sectioned on a conventional cryostat. Semithin sections (2-4 microns) alternatingly stained for parvalbumin and glutamate decarboxylase enabled us to demonstrate the coexistence of both antigens in the same cell. Thick sections (40 microns) of central and peripheral nervous system tissue were immunostained and processed for correlated light and electron microscopic studies. At the electron microscopic level, the preservation of ultrastructural features such as membranes and synaptic contacts was comparable to that normally seen in vibratome sectioned material. Hence, this technique can successfully be used for preembedding coexistence studies and electron microscopic preembedding immunocytochemistry when vibratome sectioning is problematic.
灌注固定的组织块在高摩尔蔗糖溶液中孵育,在融化的异戊烷中速冻,并在传统低温恒温器上切片。对小清蛋白和谷氨酸脱羧酶进行交替染色的半薄切片(2 - 4微米)使我们能够证明两种抗原在同一细胞中共存。对中枢和外周神经系统组织的厚切片(40微米)进行免疫染色,并进行相关的光镜和电镜研究。在电子显微镜水平上,膜和突触接触等超微结构特征的保存与振动切片机切片材料中通常所见的相当。因此,当振动切片有问题时,该技术可成功用于包埋前共存研究和电子显微镜包埋前免疫细胞化学。