■改变的纤维蛋白纤维结构与病理状态有关,包括冠心病,缺血性卒中,和动脉粥样硬化。然而,几种不同的技术通常用于研究纤维蛋白结构,并且由于缺乏标准化,使用不同技术获得的结果的比较可能具有挑战性。
■该研究通过使用多种不同的互补实验方法比较一系列生理纤维蛋白原和凝血酶浓度的纤维蛋白纤维直径,从而提供了标准化的途径。
■我们使用扫描电子显微镜(SEM)测定纤维直径,超分辨率(随机光学重建显微镜)荧光显微镜,和4通常使用比浊法来确定结果与各自应使用的条件之间的一致性。
■我们发现,对于几乎所有测试条件,使用SEM和超分辨率成像获得的直径值在10%以内。我们还发现,当使用500至800nm的波长范围进行测量并考虑折射率和特定折射率增量的波长依赖性时,在大多数情况下,使用校正的Yeromonahos浊度计方法获得的直径与SEM在20%以内。
■我们进行了系统的,评估一系列生化条件下纤维蛋白纤维直径的多技术调查。使用SEM和超分辨率成像获得的直径值的相似性表明在SEM样品制备期间的干燥和固定不会显著改变纤维横截面。一致性,在一定条件下,在使用SEM获得的直径值之间,超分辨率荧光成像,和比浊法证明了纤维蛋白直径标准化项目的可行性。
UNASSIGNED: Altered fibrin fiber structure is linked to pathologic states, including coronary heart disease, ischemic stroke, and atherosclerosis. However, several different techniques are commonly utilized for studying fibrin structures, and comparison of results obtained using different techniques can be challenging due to lack of standardization.
UNASSIGNED: This study provides a path toward standardization by comparing fibrin fiber diameters for a range of physiologic fibrinogen and thrombin concentrations using multiple different complementary experimental methods.
UNASSIGNED: We determined fiber diameter using scanning electron microscopy (SEM), superresolution (stochastic optical reconstruction microscopy) fluorescence microscopy, and 4 commonly utilized turbidimetric approaches to determine the congruence between the results and the conditions under which each should be used.
UNASSIGNED: We found that diameter values obtained using SEM and superresolution imaging agree within 10% for nearly all conditions tested. We also found that when a wavelength range of 500 to 800 nm was used for measurements and accounting for the wavelength dependence of the refractive index and specific refractive index increment, diameters obtained using the corrected Yeromonahos turbidimetric approach agree with SEM within 20% for most conditions.
UNASSIGNED: We performed a systematic, multitechnique survey assessing fibrin fiber diameters under a range of biochemical conditions. The similarity in the diameter values obtained using SEM and superresolution imaging suggests that drying and fixation during SEM sample preparation do not dramatically alter fiber cross-sections. Congruence, under certain conditions, between diameter values obtained using SEM, superresolution fluorescence imaging, and turbidimetry demonstrates the feasibility of a fibrin diameter standardization project.