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低角度光散射分析:一种用于人类和鼠类血小板受体功能特征分析的新型定量方法。

Low angle light scattering analysis: a novel quantitative method for functional characterization of human and murine platelet receptors.

机构信息

Institute of Clinical Biochemistry and Pathobiochemistry, University of Wuerzburg, Wuerzburg, Germany.

出版信息

Clin Chem Lab Med. 2012 Jul;50(7):1253-62. doi: 10.1515/CCLM.2011.817.

Abstract

BACKGROUND

Determinations of platelet receptor functions are indispensable diagnostic indicators of cardiovascular and hemostatic diseases including hereditary and acquired receptor defects and receptor responses to drugs. However, presently available techniques for assessing platelet function have some disadvantages, such as low sensitivity and the requirement of large sample sizes and unphysiologically high agonist concentrations. Our goal was to develop and initially characterize a new technique designed to quantitatively analyze platelet receptor activation and platelet function on the basis of measuring changes in low angle light scattering.

METHODS

We developed a novel technique based on low angle light scattering registering changes in light scattering at a range of different angles in platelet suspensions during activation.

RESULTS

The method proved to be highly sensitive for simultaneous real time detection of changes in size and shape of platelets during activation. Unlike commonly-used methods, the light scattering method could detect platelet shape change and aggregation in response to nanomolar concentrations of extracellular nucleotides. Furthermore, our results demonstrate that the advantages of the light scattering method make it a choice method for platelet receptor monitoring and for investigation of both murine and human platelets in disease models.

CONCLUSIONS

Our data demonstrate the suitability and superiority of this new low angle light scattering method for comprehensive analyses of platelet receptors and functions. This highly sensitive, quantitative, and online detection of essential physiological, pathophysiological and pharmacological-response properties of human and mouse platelets is a significant improvement over conventional techniques.

摘要

背景

血小板受体功能的测定是心血管和止血疾病(包括遗传性和获得性受体缺陷以及受体对药物的反应)不可或缺的诊断指标。然而,目前用于评估血小板功能的技术存在一些缺点,例如灵敏度低、需要大量样本和非生理高浓度激动剂。我们的目标是开发一种新的技术,并初步对其进行特征描述,该技术旨在基于测量低角度光散射的变化来定量分析血小板受体激活和血小板功能。

方法

我们开发了一种基于低角度光散射的新技术,该技术在血小板悬浮液激活过程中可在不同角度范围内登记光散射的变化。

结果

该方法在实时检测血小板激活过程中的大小和形状变化方面具有很高的灵敏度。与常用方法不同,光散射方法可以检测到纳米摩尔浓度的细胞外核苷酸引起的血小板形状变化和聚集。此外,我们的结果表明,光散射方法的优势使其成为监测血小板受体和研究疾病模型中鼠和人血小板的首选方法。

结论

我们的数据表明,这种新的低角度光散射方法非常适合全面分析血小板受体和功能。与传统技术相比,该方法对人血小板和鼠血小板的基本生理、病理生理和药理反应特性具有高度灵敏性、定量性和在线检测能力,是一种显著的改进。

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