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用戊二醛-十六烷基氯化吡啶固定后花粉表面物质中伴刀豆球蛋白A受体位点的电子显微镜定位

Electron microscopic localization of concanavalin A receptor sites in pollen surface material after fixation with glutaraldehyde-cetylpyridinium chloride.

作者信息

Grote M, Fromme H G

出版信息

J Histochem Cytochem. 1984 Aug;32(8):869-71. doi: 10.1177/32.8.6430993.

Abstract

Pollen from birch trees (Betula pendula) was fixed in glutaraldehyde containing 0.5% cetylpyridinium chloride (CPC), incubated with concanavalin A (Con A)-ferritin, postfixed in osmium, dehydrated, and embedded in Epon. On ultrathin sections, ferritin particles were observed closely associated with the electron-dense material precipitated by CPC on the surface of the pollen grains. Controls for CPC, which were fixed in glutaraldehyde alone, showed no electron-dense material on the surface. In controls for Con A, which were incubated in Con A-ferritin in the presence of the inhibitory sugar (alpha-methyl-D-mannopyranoside), no ferritin particles were observed. The above-described procedure thus allows the localization of sugar residues in highly soluble pollen wall glycoproteins.

摘要

桦树(垂枝桦)的花粉用含有0.5%十六烷基氯化吡啶(CPC)的戊二醛固定,与伴刀豆球蛋白A(Con A)-铁蛋白一起孵育,用锇后固定,脱水,并包埋在环氧树脂中。在超薄切片上,观察到铁蛋白颗粒与花粉粒表面由CPC沉淀的电子致密物质紧密相关。仅用戊二醛固定的CPC对照在表面未显示电子致密物质。在含有抑制性糖(α-甲基-D-甘露吡喃糖苷)的情况下用Con A-铁蛋白孵育的Con A对照未观察到铁蛋白颗粒。因此,上述方法可用于定位高度可溶的花粉壁糖蛋白中的糖残基。

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