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低温包埋胰腺组织中结构保存和免疫细胞化学染色的增强

Enhancement of structural preservation and immunocytochemical staining in low temperature embedded pancreatic tissue.

作者信息

Roth J, Bendayan M, Carlemalm E, Villiger W, Garavito M

出版信息

J Histochem Cytochem. 1981 May;29(5):663-71. doi: 10.1177/29.5.6166664.

Abstract

The recently developed low temperature embedding procedure with the resin Lowicryl K4M (Carlemalm E, Garavito M, Villiger W: Proc 7th Eur Cong Electron Microsc, 1980, p 656; Garavito M, Carlemalm E, Villiger W: Proc 7th Eur Cong Electron Microsc, 1980, p 658) was tested for its suitability for embedding of glutaraldehyde-fixed rat pancreatic tissue and for postembedding staining of thin sections with the protein A-gold (pAg) technique (Roth J, Bendayan M, Orci L: J Histochem Cytochem 26:1074, 1978) for amylase. Compared to conventional Epon embedding of glutaraldehyde fixed tissue, the low temperature embedding method with Lowicryl K4M resulted in a superior preservation of the general cellular fine structure, particularly in the Golgi apparatus. For low temperature embedded tissue, the quantitative evaluation of the immunocytochemical labeling for amylase showed a more specific staining of the rough endoplasmic reticulum, the Golgi apparatus, and the zymogen granules. This was due to a significant lowering of the background staining over all cellular organelles. The use of Lowicryl K4M at low temperature, due to the superior preservation, yields improved resolution and specificity in immunocytochemical postembedding staining.

摘要

最近开发的使用Lowicryl K4M树脂的低温包埋程序(Carlemalm E,Garavito M,Villiger W:《第七届欧洲电子显微镜大会论文集》,1980年,第656页;Garavito M,Carlemalm E,Villiger W:《第七届欧洲电子显微镜大会论文集》,1980年,第658页),已针对其对戊二醛固定的大鼠胰腺组织包埋的适用性以及用蛋白A-金(pAg)技术(Roth J,Bendayan M,Orci L:《组织化学与细胞化学杂志》26:1074,1978年)对淀粉酶进行薄切片后包埋染色的适用性进行了测试。与戊二醛固定组织的传统环氧树脂包埋相比,使用Lowicryl K4M的低温包埋方法能更好地保存一般细胞的精细结构,尤其是高尔基体。对于低温包埋的组织,淀粉酶免疫细胞化学标记的定量评估显示,粗面内质网、高尔基体和酶原颗粒的染色更具特异性。这是由于所有细胞器的背景染色显著降低。由于保存效果更好,在低温下使用Lowicryl K4M可提高免疫细胞化学后包埋染色的分辨率和特异性。

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