Cliff Ben, Lockyer Nicholas, Jungnickel Harald, Stephens Gill, Vickerman John C
Surface Analysis Research Centre, Department of Chemistry, UMIST, Manchester M60 1QD, UK.
Rapid Commun Mass Spectrom. 2003;17(19):2163-7. doi: 10.1002/rcm.1169.
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.
成像静态二次离子质谱法（SIMS）提供了一种从生物系统中获取分子信息的强大方法，具有良好的空间分辨率。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。我们在此报告一种新型冷冻断裂装置的开发，以促进对生物细胞材料的操作和分析，通过快速冷冻使细胞化学性质保持完整。我们用冷冻断裂、冷冻水合脂质体的高对比度图像来说明性能特征，脂质双层中含有氯法齐明药物作为标记物，用于确定穿过脂质体结构的断裂平面。通过监测和成像处于冷冻水合状态的酵母细胞表面的氯法齐明，并证明在用于揭示细胞超微结构的断裂细胞的分子信息中不存在该分子，我们证明该分子不会穿透细胞壁。
