Standardization and Validation of Fluorescence-Based Quantitative Assay to Study Human Platelet Adhesion to Extracellular-Matrix in a 384-Well Plate

基于荧光的定量分析的标准化和验证,用于研究 384 孔板中人血小板对细胞外基质的粘附性

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作者:Augusto Martins Lima, Damian S Saint Auguste, François Cuenot, Ana C Martins Cavaco, Tom Lachkar, Cindy Marie Elodie Khawand, Rodrigo A Fraga-Silva, Nikolaos Stergiopulos

Abstract

Platelets play a crucial role in the immunological response and are involved in the pathological settings of vascular diseases, and their adhesion to the extracellular matrix is important to bring leukocytes close to the endothelial cells and to form and stabilize the thrombus. Currently there are several methods to study platelet adhesion; however, the optimal parameters to perform the assay vary among studies, which hinders their comparison and reproducibility. Here, a standardization and validation of a fluorescence-based quantitative adhesion assay to study platelet-ECM interaction in a high-throughput screening format is proposed. Our study confirms that fluorescence-based quantitative assays can be effectively used to detect platelet adhesion, in which BCECF-AM presents the highest sensitivity in comparison to other dyes.

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