1.1 本指南为二次离子质谱（SIMS）分析员提供了分析体外生长的单个组织培养细胞的低温方法。本指南适用于冷冻水合和冷冻干燥样品类型。包括相关光学、激光扫描共焦和二次电子显微镜的程序，以补充SIMS分析。 1.2 本指南不适用于不附着在基质上的细胞培养。 1.3 本指南不适用于任何塑料嵌入细胞培养标本。 1.4 本标准并非旨在解决与其使用相关的所有安全问题（如有）。本标准的用户有责任在使用前制定适当的安全、健康和环境实践，并确定监管限制的适用性。 1.5 本国际标准是根据世界贸易组织技术性贸易壁垒（TBT）委员会发布的《关于制定国际标准、指南和建议的原则的决定》中确立的国际公认标准化原则制定的。 ====意义和用途====== 5.1 细胞生长培养基的存在使用二甲基硅氧烷直接分析细胞变得复杂。试图洗掉营养培养基会导致细胞暴露于非生理试剂中，这也可能改变其化学成分。通过使用夹层冻结断裂方法克服了这一障碍 ( 1. ) . 这种低温方法提供了一种独特的方法，可以在原始状态下对单个电池进行采样，以进行二次离子质谱分析。 5.2 本文描述的程序已成功用于Na成像 + 和K + 离子输运 ( 3. ) ，受刺激细胞中的钙改变 ( 4. , 5. ) ，以及治疗药物和同位素标记分子在单细胞中的定位 ( 6. ) . 已检查根据该方法制备的冷冻冻干细胞的SIMS基质效应 ( 7. ) . 在这种样品类型中也实现了离子图像量化 ( 8. ) . 5.3 本文描述的程序适用于多种细胞培养，并为研究单个细胞对健康和疾病状态下化学变化的反应以及同位素定位提供了一种方法- 细胞培养模型中的标记分子和治疗药物。

1.1 This guide provides the Secondary Ion Mass Spectrometry (SIMS) analyst with a cryogenic method for analyzing individual tissue culture cells growing in vitro. This guide is suitable for frozen-hydrated and frozen-freeze-dried sample types. Included are procedures for correlating optical, laser scanning confocal and secondary electron microscopies to complement SIMS analysis. 1.2 This guide is not suitable for cell cultures that do not attach to the substrate. 1.3 This guide is not suitable for any plastic embedded cell culture specimens. 1.4 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use. 1.5 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee. ====== Significance And Use ====== 5.1 The presence of cell growth medium complicates a direct analysis of cells with SIMS. Attempts to wash out the nutrient medium results in the exposure of cells to unphysiological reagents that may also alter their chemical composition. This obstacle is overcome by using a sandwich freeze-fracture method ( 1 ) . This cryogenic method has provided a unique way of sampling individual cells in their native state for SIMS analysis. 5.2 The procedure described here has been successfully used for imaging Na + and K + ion transport ( 3 ) , calcium alterations in stimulated cells ( 4 , 5 ) , and localization of therapeutic drugs and isotopically labeled molecules in single cells ( 6 ) . The frozen freeze-dried cells prepared according to this method have been checked for SIMS matrix effects ( 7 ) . Ion image quantification has also been achieved in this sample type ( 8 ) . 5.3 The procedure described here is amenable to a wide variety of cell cultures and provides a way for studying the response of individual cells for chemical alterations in the state of health and disease and localization of isotopically-labeled molecules and theraputic drugs in cell culture models.