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“远程自旋捕获”:一种扩大自旋捕获测量可用性的方法。

"Distant spin trapping": a method for expanding the availability of spin trapping measurements.

作者信息

Khan Nadeem, Grinberg Oleg, Wilmot Carmen, Kiefer Heather, Swartz Harold M

机构信息

Department of Diagnostic Radiology, EPR Center for the Study of Viable Systems, 7785 Vail, Room 702, Dartmouth Medical School, Hanover, NH 03755, USA.

出版信息

J Biochem Biophys Methods. 2005 Feb 28;62(2):125-30. doi: 10.1016/j.jbbm.2004.10.001. Epub 2004 Nov 13.

Abstract

The technique of spin trapping is used to study a wide range of free radicals in various systems, including those generated in vitro and in vivo. But unfortunately, EPR spectrometers are not always immediately accessible at the site of experimentation, and therefore it is important to find a method that can preserve a radical adduct over longer periods of time. We describe here an alternative method in which the samples can be frozen and transported for EPR measurements at another site. Various spin adducts of DEPMPO were frozen and measured at 0 degrees C at various intervals after freezing to determine their stability in the frozen state. The radical adducts were generated by established methods and stored at two different temperatures; -196 degrees C (liquid nitrogen) and -80 degrees C (dry ice). The experiments were carried out in an aqueous solution with and without a model of reducing environment (2 mM ascorbate). The results indicate that it is feasible to store and transport spin adducts for subsequent analysis. We conclude that this approach, which we term "distant spin trapping", makes it feasible to transport samples to another site for EPR measurements. This should significantly expand the ability to use spin trapping in biology and medicine.

摘要

自旋捕集技术用于研究各种体系中的多种自由基,包括体外和体内产生的自由基。但不幸的是,在实验现场并非总能立即使用电子顺磁共振(EPR)光谱仪,因此找到一种能在较长时间内保存自由基加合物的方法很重要。我们在此描述一种替代方法,即可以将样品冷冻并运至另一地点进行EPR测量。将DEPMPO的各种自旋加合物冷冻,并在冷冻后的不同时间间隔于0℃下进行测量,以确定它们在冷冻状态下的稳定性。自由基加合物通过既定方法生成,并保存在两种不同温度下:-196℃(液氮)和-80℃(干冰)。实验在有和没有还原环境模型(2 mM抗坏血酸盐)的水溶液中进行。结果表明,储存和运输自旋加合物以供后续分析是可行的。我们得出结论,这种我们称为“远程自旋捕集”的方法使将样品运至另一地点进行EPR测量成为可能。这将显著扩展在生物学和医学中使用自旋捕集的能力。

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