Improved measurement of the rotor temperature in analytical ultracentrifugation
Improved measurement of the rotor temperature in analytical ultracentrifugation
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DOI：
10.1016/j.ab.2014.02.006
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发表时间：
2014-04-15
影响因子：
2.9
通讯作者：
Schuck, Peter
中科院分区：
文献类型：
--
作者：
Zhao, Huaying;Balbo, Andrea;Schuck, Peter
沉降速度是测量生物大分子流体动力学、平移摩擦系数的经典方法。在最近的一项研究中，我们比较了各种分析超分辨率，我们表明扫描时间，径向放大率和温度的外部校准对于准确测量至关重要（Anal. 440（2013）81-95）。为了实现精确的温度校准，我们引入了一种自主微型温度记录集成电路（Maxim Thermochron iButton），该集成电路可插入超离心电池组件并以低转子速度旋转。在目前的工作中，我们开发了一种改进的保持器，用于位于转子手柄中的温度传感器。这具有不降低转子容量的优点，并且允许在高达60，000 rpm的高速沉降速度实验期间直接测量纺丝转子的温度。我们证明了这种方法的灵敏度，通过监测绝热冷却由于转子拉伸在转子加速和转子减速的反向过程。在此基础上，我们开发了一个程序，以近似等温转子加速更好的温度控制。爱思唯尔公司出版
Sedimentation velocity is a classical method for measuring the hydrodynamic, translational friction coefficient of biological macromolecules. In a recent study comparing various analytical ultracentrifuges, we showed that external calibration of the scan time, radial magnification, and temperature is critically important for accurate measurements (Anal. Biochem. 440 (2013) 81-95). To achieve accurate temperature calibration, we introduced the use of an autonomous miniature temperature logging integrated circuit (Maxim Thermochron iButton) that can be inserted into an ultracentrifugation cell assembly and spun at low rotor speeds. In the current work, we developed an improved holder for the temperature sensor located in the rotor handle. This has the advantage of not reducing the rotor capacity and allowing for a direct temperature measurement of the spinning rotor during high-speed sedimentation velocity experiments up to 60,000 rpm. We demonstrated the sensitivity of this approach by monitoring the adiabatic cooling due to rotor stretching during rotor acceleration and the reverse process on rotor deceleration. Based on this, we developed a procedure to approximate isothermal rotor acceleration for better temperature control. Published by Elsevier Inc.