Abstract Number: PB1699
Meeting: ISTH 2020 Congress
Background: Although parallel-plate flow chamber is a widely used tool for whole blood arterial thrombus formation studies and has potential for diagnostic use, the details of thrombus dynamics and structure, spatial distribution in the chamber, pre-analytical and analytical conditions, and proper quantification methods are not clear.
Aims: To investigate parameters of thrombus formation induced by a collagen spot in a parallel-plate flow chamber.
Methods: Hirudinated whole blood was pumped at a constant velocity in a 50 um height parallel plate flow chamber to produce shear rate of 1000 s-1. DiOC6 and anti-P-selectin antibodies were used for labeling. Activation was by 1×1 mm collagen-coated spot. Spinning-disc confocal and epifluorescent videomicroscopy were used for imaging.
Results: Thrombi volume increased linearly for 10 minutes as observed by fluorescent intensity of DiOC6, followed by disruption and embolization. Local maximum of DiOC6 fluorescence was proportional to thrombi height, with the maximal height reached of about 40% of the channel height. Surface coverage increased as a sigmoid function of time and eventually reached the plateau value of 60% on average. P-selectin staining was present in the thrombi base but did not reach the tops of the thrombi. Although surface coverage was uniform throughout the collagen-covered spot, thrombi volume and height decreased downstream, with the largest thrombi located at the front border of the collagen-coated spot. Comparison of automatic image analysis algorithms has shown that fixed global thresholding applied to fluorescent images gives inaccurate results in determining surface coverage that can be improved using a density-based algorithm.
Conclusions: Thrombus formation in this flow chambers is characterized by heterogeneity and embolization.
To cite this abstract in AMA style:Ryabykh A, Beresneva E, Nechipurenko D, Panteleev M. Characterization of Thrombus Heterogeneity and Dynamics of Formation on Collagen Spots in a Parallel-plate Flow Chamber [abstract]. Res Pract Thromb Haemost. 2020; 4 (Suppl 1). https://abstracts.isth.org/abstract/characterization-of-thrombus-heterogeneity-and-dynamics-of-formation-on-collagen-spots-in-a-parallel-plate-flow-chamber/. Accessed January 23, 2022.
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