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使用40 keV C60+ 初级离子束对脑组织切片进行深度剖析。

Depth profiling brain tissue sections with a 40 keV C60+ primary ion beam.

作者信息

Jones Emrys A, Lockyer Nicholas P, Vickerman John C

机构信息

Surface Analysis Research Centre, Manchester Interdisciplinary Biocentre, The University of Manchester, 131 Princess Street, Manchester, M1 7DN, United Kingdom.

出版信息

Anal Chem. 2008 Mar 15;80(6):2125-32. doi: 10.1021/ac702127q. Epub 2008 Feb 16.

DOI:10.1021/ac702127q
PMID:18278949
Abstract

In this paper, the effect of prolonged C60(+) primary ion bombardment on the chemical information available from a section of rat brain is discussed. Initial attempts demonstrate the rapid loss of molecular signal from the bombarded area with both C60(+) and Au(+) used as a monatomic comparison. However, the nature of this signal disappearance is shown to be different. Analysis of the C60(+) data indicates a correlation between signal loss and the appearance of sodium and potassium adducts of phosphate and protein fragments; this is supported by model systems. By using an ammonium formate wash to reduce the salt levels within the tissue this effect is removed, allowing the chemistry of the tissue section to be better probed. Results collected from multiple sections suggest that at room temperature under vacuum conditions there is a migration of lipids to the surface of the tissue. Three-dimensional (3D) imaging is used to demonstrate that once these lipids are removed other species, such as proteins, are uncovered. By depth profiling the sample in a frozen state, the degree and importance of lipid migration to the observed localization of native compounds is assessed. This investigation into the behavior of biological tissue under high C60(+) fluxes not only allows an evaluation of the potential accuracy of 3D SIMS mapping of important biological molecules but also demonstrates the possibility of using ion doses beyond the traditional "static limit" to provide higher secondary ion yields that could lead to greater detection limits and smaller useful lateral resolution within such analyses.

摘要

本文讨论了长时间的C60(+) 初级离子轰击对大鼠脑切片中可用化学信息的影响。初步尝试表明,以C60(+) 和用作单原子对照的Au(+) 进行轰击时,轰击区域的分子信号会迅速丧失。然而,这种信号消失的性质被证明是不同的。对C60(+) 数据的分析表明,信号损失与磷酸盐和蛋白质片段的钠和钾加合物的出现之间存在相关性;模型系统支持了这一点。通过使用甲酸铵洗涤来降低组织内的盐水平,这种影响得以消除,从而能够更好地探测组织切片的化学性质。从多个切片收集的结果表明,在真空条件下的室温下,脂质会迁移到组织表面。三维 (3D) 成像用于证明一旦去除这些脂质,其他物质(如蛋白质)就会暴露出来。通过对冷冻状态下的样品进行深度剖析,评估了脂质迁移对天然化合物观察到的定位的程度和重要性。这项对高C60(+) 通量下生物组织行为的研究不仅有助于评估重要生物分子的3D二次离子质谱 (SIMS) 映射的潜在准确性,还证明了使用超出传统 “静态极限” 的离子剂量以提供更高的二次离子产率的可能性,这可能会导致在此类分析中获得更高的检测限和更小的有效横向分辨率。

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