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首页> 外文期刊>Applied Surface Science >Cations in mammalian cells and chromosomes: Sample preparation protocols affect elemental abundances by SIMS
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Cations in mammalian cells and chromosomes: Sample preparation protocols affect elemental abundances by SIMS

机译：哺乳动物细胞和染色体中的阳离子：样品制备方案会通过SIMS影响元素丰度

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The focus of our current research aims at detailing and quantifying the presence of cations, primarily Ca and Mg, in mammalian cells and chromosomes throughout the different stages of the cell cycle, using our high resolution scanning ion microprobe, the UGSIM. The 45 keV Ga+ probe of this instrument, typically similar to 40 nm in diameter, carries a current of 30-40 pA, appropriate for surface SIMS studies, but limited in sample erosion rate for dynamic SIMS mapping over cell-size areas, of order 100 mu m x 100 mu m. Practical and reliable use of this probe toward the above SIMS goals requires a careful matching of the latter factors with the physical and chemical consequences of sample preparation protocols. We examine here how the preferred sample cryo-preservation methodologies such as freeze-fracture and lyophilization affect high resolution SIMS analysis, and, from this standpoint, develop and evaluate the advantages and disadvantages of fast alternate approaches to drying frozen samples. The latter include the use of methanol, ethanol, and methanol/acetic acid fixative. Methanol-dried freeze-fractured samples preserve histological morphology and yield Ca and Mg distributions containing reliable differential dynamical information, when compared with those following lyophilization. (c) 2006 Elsevier B.V. All rights reserved.
机译：我们目前的研究重点是使用高分辨率扫描离子微探针UGSIM详细描述和量化哺乳动物细胞和染色体在整个细胞周期各个阶段中阳离子（主要是Ca和Mg）的存在。该仪器的45 keV Ga +探针通常直径类似于40 nm，载有30-40 pA的电流，适用于表面SIMS研究，但对于在细胞大小区域上进行动态SIMS映射的样品腐蚀速率有所限制100微米x 100微米为了达到上述SIMS目标，要切实可行地使用此探针，需要将后一因素与样品制备规程的物理和化学后果进行仔细匹配。我们在这里检查首选的样品冷冻保存方法，例如冷冻破裂和冻干方法如何影响高分辨率SIMS分析，并从这个角度出发，开发和评估快速替代方法干燥冷冻样品的优点和缺点。后者包括使用甲醇，乙醇和甲醇/乙酸固定剂。与冷冻干燥后的样品相比，经甲醇干燥的冷冻断裂样品保留了组织学形态，并产生了包含可靠的差分动力学信息的Ca和Mg分布。 （c）2006 Elsevier B.V.保留所有权利。

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