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使用动态光散射测定水溶液 DNA 溶液的黏度。

Viscosity of aqueous DNA solutions determined using dynamic light scattering.

机构信息

Department of Chemistry and Warwick Centre for Analytical Science, University of Warwick, Coventry, CV4 7AL, UK.

出版信息

Analyst. 2011 Oct 21;136(20):4159-63. doi: 10.1039/c1an15475c. Epub 2011 Aug 25.

DOI:10.1039/c1an15475c
PMID:21869949
Abstract

Viscosity is a key parameter for characterising the behaviour of liquids and their flow. It is, however, difficult to measure precisely, reproducibly and accurately for aqueous solutions on a micro-litre volume scale, which is what is usually needed for biological samples. We report the development of a new method for measuring dynamic viscosity by measuring dynamic light scattering (DLS) data for a range of particles of well-defined size. Most applications of DLS involve determining particle size for samples of known viscosity. We inverted the usual protocol and endeavoured to determine viscosity for samples of known particle size. Viscosity measurements for water and aqueous solutions of calf thymus DNA made using DLS were compared with those from a U-tube viscometer. The styrene particles, frequently used as particle size standards, gave unsatisfactory results for our DNA samples as did C-6 derivatized silica and positively charged amino polystyrene microspheres. However, negatively charged carboxylate polystyrene microspheres particles readily gave accurate viscosity measurements over a range of temperatures (0-100 °C). The sample volume required depends on the cuvette used to measure DLS, but can be performed with samples sizes ranging from 40 to 3000 μL. The sample can then be recovered for subsequent experiments. The DLS method is simple to perform at different temperatures and provides data of accuracy significantly above that of a U-tube viscometer. Our results also indicate a way forward to account accurately for solution viscosity in the normal applications of DLS to particle size determination by including the appropriate non-interacting particles as an internal standard.

摘要

粘度是表征液体及其流动行为的关键参数。然而,在微升体积范围内,很难准确、可重复和精确地测量水溶液的粘度,而这正是生物样品通常需要的。我们报告了一种通过测量一系列具有明确定义大小的粒子的动态光散射 (DLS) 数据来测量动态粘度的新方法。DLS 的大多数应用涉及确定具有已知粘度的样品的粒径。我们反转了通常的方案,并努力确定具有已知粒径的样品的粘度。使用 DLS 对水和小牛胸腺 DNA 水溶液进行的粘度测量与来自 U 型管粘度计的测量结果进行了比较。对于我们的 DNA 样品,苯乙烯粒子(通常用作粒径标准)以及 C-6 衍生化的硅和带正电荷的氨基聚苯乙烯微球的结果并不令人满意。然而,带负电荷的羧酸盐聚苯乙烯微球粒子在 0-100°C 的范围内很容易给出准确的粘度测量值。所需的样品体积取决于用于测量 DLS 的比色皿,但可以使用 40 至 3000 μL 的样品体积进行测量。然后可以回收样品用于后续实验。DLS 方法在不同温度下简单易行,并提供了明显高于 U 型管粘度计的精度的数据。我们的结果还表明了一种方法,可以通过包括适当的非相互作用粒子作为内标,准确地考虑溶液粘度在 DLS 正常应用于粒径测定中的作用。

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