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用于神经组织光镜免疫细胞化学的锌-醛固定法。

Zinc-aldehyde fixation for light-microscopic immunocytochemistry of nervous tissues.

作者信息

Mugnaini E, Dahl A L

出版信息

J Histochem Cytochem. 1983 Dec;31(12):1435-8. doi: 10.1177/31.12.6355290.

Abstract

Two procedures are described for vascular perfusion of the nervous system with a zinc-aldehyde fixative. The procedures, simple and economical, combine the advantages of perfusion fixation with an aldehyde solution and matrix stabilization by a mordating agent, and improve the sensitivity of the peroxidase-antiperoxidase (PAP) method for the immunocytochemical localization of several antigens. Procedure A is intended for the light-microscopic immunostaining of cellular elements containing high concentrations of antigen. Penetration of the immunoreagents is adequate without the use of detergents. Procedure B is particularly advantageous for the light-microscopic immunostaining of cellular elements that contain low concentrations of antigen, and for high-resolution microphotography. With procedure B, the tissue penetration of immunoreagents is more limited than with procedure A; however, neuronal cell bodies and dendrites are more easily penetrated by the immunoreagents than are axons. Neuronal cell bodies and dendrites thus become clearly detectable in the light-microscope, even when they are surrounded by numerous immunoreactive axon terminals, and especially after the blockage of axoplasmic transport by the topical injection of colchicine.

摘要

本文描述了两种用锌-醛固定剂对神经系统进行血管灌注的方法。这些方法简单且经济,结合了醛溶液灌注固定和媒染剂稳定基质的优点,并提高了过氧化物酶-抗过氧化物酶(PAP)法对几种抗原进行免疫细胞化学定位的敏感性。方法A适用于对含有高浓度抗原的细胞成分进行光学显微镜免疫染色。无需使用去污剂,免疫试剂就能充分渗透。方法B对于含有低浓度抗原的细胞成分的光学显微镜免疫染色以及高分辨率显微摄影特别有利。采用方法B时,免疫试剂在组织中的渗透比方法A更有限;然而,免疫试剂更容易穿透神经元细胞体和树突,而不是轴突。因此,即使神经元细胞体和树突被大量免疫反应性轴突终末包围,在光学显微镜下也能清晰地检测到它们,尤其是在局部注射秋水仙碱阻断轴浆运输之后。

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