Fan Natalie K, Keegan Philip M, Platt Manu O, Averett Rodney D
Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology & Emory University School of Medicine.
Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology & Emory University School of Medicine; Parker H. Petit Institute for Bioengineering & Bioscience, Georgia Institute of Technology.
J Vis Exp. 2015 Apr 1(98):e52019. doi: 10.3791/52019.
Fibrin is an extracellular matrix protein that is responsible for maintaining the structural integrity of blood clots. Much research has been done on fibrin in the past years to include the investigation of synthesis, structure-function, and lysis of clots. However, there is still much unknown about the morphological and structural features of clots that ensue from patients with disease. In this research study, experimental techniques are presented that allow for the examination of morphological differences of abnormal clot structures due to diseased states such as diabetes and sickle cell anemia. Our study focuses on the preparation and evaluation of fibrin clots in order to assess morphological differences using various experimental assays and confocal microscopy. In addition, a method is also described that allows for continuous, real-time calculation of lysis rates in fibrin clots. The techniques described herein are important for researchers and clinicians seeking to elucidate comorbid thrombotic pathologies such as myocardial infarctions, ischemic heart disease, and strokes in patients with diabetes or sickle cell disease.
纤维蛋白是一种细胞外基质蛋白,负责维持血凝块的结构完整性。在过去几年中,针对纤维蛋白开展了大量研究,包括对其合成、结构功能以及血凝块溶解的研究。然而,对于患有疾病的患者所形成的血凝块的形态和结构特征,仍有许多未知之处。在本研究中,介绍了一些实验技术,这些技术可用于检查因糖尿病和镰状细胞贫血等疾病状态导致的异常血凝块结构的形态差异。我们的研究重点是纤维蛋白凝块的制备和评估,以便使用各种实验测定方法和共聚焦显微镜来评估形态差异。此外,还描述了一种方法,该方法可对纤维蛋白凝块的溶解速率进行连续、实时计算。本文所述技术对于试图阐明糖尿病或镰状细胞病患者中诸如心肌梗死、缺血性心脏病和中风等合并血栓形成病理情况的研究人员和临床医生而言非常重要。