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使用喹硫平651对半薄切片上的γ-氨基丁酸进行包埋后免疫组织化学显示。

The use of Quetol 651 for the post-embedding immunohistochemical demonstration of gamma-aminobutyric acid on semithin sections.

作者信息

Denizot J P, Arnold G, Geffard M, Libouban S

机构信息

Laboratoire de Physiologie Nerveuse, Dept. Neurophysiologie Sensorielle, CNRS, Gif-sur-Yvette, France.

出版信息

Histochem J. 1988 Apr;20(4):222-9. doi: 10.1007/BF01747467.

Abstract

Quetol 651 was used as an embedding medium for the demonstration of gamma aminobutyric acid (GABA) in semithin sections by the peroxidase-anti-peroxidase method. In order to demonstrate the immunoreactivity, the embedding medium was partially dissolved using absolute ethanol containing 0.8-1 M NaOH or KOH for 5-7 min. The experimental procedure was elaborated by testing the GABAergic sites in the endings surrounding the small neurones of the anterior exterolateral nucleus of a mormyrid fish and in the pyramidal cells of the electrosensory lateral line lobe of gymnotoid fish by applying anti-GAD (glutamic acid decarboxylase) antiserum. To test the general validity of the use of Quetol 651, GABAergic sites were also identified in the central nervous system of an insect, the honey bee, with anti-GABA and anti-GAD antisera. The intensity of labelling revealed by immunoperoxidase applied to Quetol 651-embedded semithin sections, demonstrated high precision and gave good resolution for light microscopical observations.

摘要

喹托651用作包埋介质,通过过氧化物酶-抗过氧化物酶法在半薄切片中显示γ-氨基丁酸(GABA)。为了显示免疫反应性,使用含0.8 - 1M氢氧化钠或氢氧化钾的无水乙醇将包埋介质部分溶解5 - 7分钟。通过应用抗谷氨酸脱羧酶(GAD)抗血清,检测了裸臀鱼前外侧核小神经元周围末梢以及电鱼电感觉侧线叶锥体细胞中的GABA能位点,从而详细阐述了实验过程。为了测试使用喹托651的普遍有效性,还用抗GABA和抗GAD抗血清在昆虫蜜蜂的中枢神经系统中鉴定了GABA能位点。应用于喹托651包埋的半薄切片的免疫过氧化物酶显示的标记强度,具有高精度,并且为光学显微镜观察提供了良好的分辨率。

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