Atomic and molecular imaging at the single-cell level with TOF-SIMS
Atomic and molecular imaging at the single-cell level with TOF-SIMS
复制标题
利用 TOF-SIMS 在单细胞水平上进行原子和分子成像
DOI：
10.1021/ac9701748
复制
发表时间：
1997-07-01
影响因子：
7.4
通讯作者：
Winograd, N
中科院分区：
文献类型：
--
作者：
Colliver, TL;Brummel, CL;Winograd, N
一个完整的冷链冷冻断裂方法已被开发，以测试使用飞行时间二次离子质谱（TOF-SIMS）成像的冷冻水合生物样品的分子分析的可行性。由于该技术仅对样品的前几个单层进行采样，因此样品表面的水可能是干扰的主要来源。通过将冷阱（断裂刀和外壳在-196摄氏度）放置在保持在较暖温度（-97至-113摄氏度）的断裂样品附近，可以最大限度地减少这个问题。这导致去除表面水并防止表面上的冷凝。虽然这种方法是有效的，但已经发现，由于其他基质分子的挥发性和干燥过程中赋予细胞表面的形态学效应，需要仔细控制样品升温。通过利用上述处理技术，已经可以首次证明TOF-SIMS成像技术可以用于利用亚微米离子探针束获得细胞表面上的分子种类的图像。已经用单细胞生物草履虫的TOF-SIMS获得了小烃和故意添加的掺杂剂DMSO和可卡因的图像。
A complete cold chain freeze-fracture methodology has been developed to test the feasibility of using time-of-flight secondary ion mass spectrometry (TOF-SIMS) imaging for the molecular analysis of frozen hydrated biological samples. Because the technique only samples the first few monolayers of a sample, water on the surface of a sample can be a major source of interference. This problem can be minimized by placing a cold trap (fracture knife and housing at -196 degrees C) near the fractured sample that is held at a warmer temperature (-97 to -113 degrees C). This results in removal of surface water and prevents condensation on the surface. Although this approach is effective, it has been found that sample warming needs to be carefully controlled due to the volatility of other matrix molecules and the morphological effects imparted onto the cell surface during drying. By utilizing the above handling technique, it has been possible to demonstrate for the first time that TOF-SIMS imaging technology can be used to obtain images of molecular species across a cell surface with a submicrometer ion probe beam. images of small hydrocarbons and the deliberately added dopants DMSO and cocaine have been obtained with TOF-SIMS of the single-cell organism Paramecium.