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液相传输电子显微镜原位镍染色法用于脂质体成像。

In Situ Ni Stain for Liposome Imaging by Liquid-Cell Transmission Electron Microscopy.

机构信息

Department of Chemistry, Northwestern University, Evanston, Illinois 60208, United States.

International Institute for Nanotechnology, Evanston, Illinois 60208, United States.

出版信息

Nano Lett. 2020 Jun 10;20(6):4292-4297. doi: 10.1021/acs.nanolett.0c00898. Epub 2020 May 26.

Abstract

Solvated soft matter, both biological and synthetic, can now be imaged in liquids using liquid-cell transmission electron microscopy (LCTEM). However, such systems are usually composed solely of organic molecules (low elements) producing low contrast in TEM, especially within thick liquid films. We aimed to visualize liposomes by LCTEM rather than requiring cryogenic TEM (cryoTEM). This is achieved here by imaging in the presence of aqueous metal salt solutions. The increase in scattering cross-section by the cation gives a staining effect that develops , which could be captured by real space TEM and verified by energy dispersive x-ray spectroscopy (EDS). We identified beam-induced staining as a time-dependent process that enhances contrast to otherwise low contrast materials. We describe the development of this imaging method and identify conditions leading to exceptionally low electron doses for morphology visualization of unilamellar vesicles before beam-induced damage propagates.

摘要

溶剂化软物质,无论是生物的还是合成的,现在都可以使用液相透射电子显微镜(LCTEM)在液体中成像。然而,这样的系统通常仅由有机分子(低 Z 元素)组成,在 TEM 中产生低对比度,特别是在厚的液体膜中。我们的目的是通过 LCTEM 而不是需要低温 TEM(cryoTEM)来可视化脂质体。通过在含有水性金属盐溶液的情况下进行成像,可以实现这一点。阳离子增加的散射截面产生染色效果,这种效果会随着时间的推移而发展,可以通过实空间 TEM 捕获,并通过能量色散 X 射线光谱(EDS)验证。我们确定了束诱导染色是一种时间依赖性过程,它增强了对比度,否则对比度低的材料就会变得低对比度。我们描述了这种成像方法的发展,并确定了在束诱导损伤传播之前,用于单室囊泡形态可视化的条件,这些条件导致了异常低的电子剂量。

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