Cell-surface receptors and proteins on platelet membranes imaged by scanning force microscopy using immunogold contrast enhancement.
Cell-surface receptors and proteins on platelet membranes imaged by scanning force microscopy using immunogold contrast enhancement.
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使用免疫金对比度增强扫描力显微镜对血小板膜上的细胞表面受体和蛋白质进行成像。
DOI：
10.1016/s0006-3495(95)80228-6
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Marchant,RE
中科院分区：
文献类型：
--
作者：
Eppell,SJ;Simmons,SR;Albrecht,RM;Marchant,RE
高分辨率扫描力显微镜（SFM）图像的纤维蛋白原暴露的血小板膜。使用超锋利的碳尖端，我们能够获得膜表面特征的亚分子尺度分辨率。使用低电压，高分辨率扫描电子显微镜（LVHRSEM）对SFM中看到的相同蛋白质分子进行成像，可以证实SFM结果。我们通过SFM获得了精确的高度尺寸，并辅以LVHRSEM获得了精确的横向尺寸。使用14- nm和5-nm的金标来识别特定的膜结合生物分子，并与SFM提供对比度增强，作为观察未标记材料的有用辅助。结果表明，这些标记对于定位血小板膜表面的特定蛋白质分子和使用SFM评估这些分子的分布是有用的。14纳米的标签在膜波纹上可见，而5纳米的标签似乎很难用目前的SFM仪器配置来分辨。当使用5nm标签时，附带使用LVHRSEM可以在亚分子分辨率下检查SFM图像，并在SFM实验完成后将功能与成像的结构联系起来。
High resolution scanning force microscope (SFM) images of fibrinogen-exposed platelet membranes are presented. Using ultrasharp carbon tips, we are able to obtain submolecular scale resolution of membrane surface features. Corroboration of SFM results is achieved using low voltage, high resolution scanning electron microscopy (LVHRSEM) to image the same protein molecule that is seen in the SFM. We obtain accurate height dimensions by SFM complemented by accurate lateral dimensions obtained by LVHRSEM. The use of 14- and 5-nm gold labels to identify specific membrane-bound biomolecules and to provide contrast enhancement with the SFM is explored as a useful adjunct to observation of unlabeled material. It is shown that the labels are useful for locating specific protein molecules on platelet membrane surfaces and for assessing the distribution of these molecules using the SFM. Fourteen nm labels are shown to be visible over the membrane corrugation, whereas 5-nm labels appear difficult to resolve using the present SFM instrumental configuration. When using the 5-nm labels, collateral use of LVHRSEM allows one to examine SFM images at submolecular resolution and associate function with the structures imaged after the SFM experiment is completed.