Probing cell chemistry with time-of-flight secondary ion mass spectrometry: development and exploitation of instrumentation for studies of frozen-hydrated biological material
Probing cell chemistry with time-of-flight secondary ion mass spectrometry: development and exploitation of instrumentation for studies of frozen-hydrated biological material
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DOI：
10.1002/rcm.1169
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发表时间：
2003-01-01
影响因子：
2
通讯作者：
Vickerman, JC
中科院分区：
文献类型：
--
作者：
Cliff, B;Lockyer, N;Vickerman, JC
成像静态二次离子质谱（SIMS）提供了一种从生物系统中获取具有良好空间分辨率的分子信息的有力方法。然而，该技术需要进一步发展，以使其适用于细胞的常规分析。我们在此报道一种新型冷冻断裂装置的研发，它有助于生物细胞材料的操作和分析，通过快速冷冻使细胞化学保持完整。我们用冷冻断裂、冷冻含水脂质体的高对比度图像来说明其性能特征，药物氯法齐明被限制在脂质双层内，为确定脂质体结构的断裂面提供了一个标记。通过对冷冻含水状态下酵母细胞表面的氯法齐明进行监测和成像，并证明在揭示细胞超微结构的细胞断裂所产生的分子信息中不存在氯法齐明，我们证明该分子不会穿透细胞壁。版权所有（C）2003约翰威立父子有限公司
Imaging static secondary ion mass spectrometry (SIMS) offers a powerful method of obtaining molecular information from biological systems with good spatial resolution. However, the technique needs further development to make it suitable for routine analysis of cells. We report here the development of a new freeze-facture device to facilitate the manipulation and analysis of biological cell material, with the cell chemistry preserved intact by rapid freezing. We illustrate performance characteristics with high-contrast images of freeze-fractured, frozen-hydrated liposomes with the drug clofazamine constrained within the lipid bilayer providing a marker to determine the fracture plane across the liposome structure. By monitoring and imaging clofazamine on the surface of yeast cells in the frozen-hydrated state, and demonstrating its absence within molecular information from a cell fractured to reveal the cell ultrastructure, we demonstrate that the molecule does not penetrate the cell wall. Copyright (C) 2003 John Wiley Sons, Ltd.