It is widely recognized that large platelets are generally more reactive than small platelets. Epidemiological studies have identified a link between larger platelet size and an increased risk of thrombotic complications. However, the functional differences between large and small platelets remain poorly understood, largely due to the absence of standardized protocols for size-based platelet separation. The aim of the current master thesis is to investigate the functional and morphological differences between small and large platelets by separating them into distinct fractions and studying their activation on collagen-coated surfaces. The study visualized and analyzed P-Selectin (CD62P) expression in both platelet fractions during activation-induced shape and functional changes. Using an activation protocol for photoswitching CD62P conjugated Alexa Fluor© 647 we successfully captured super-resolution 3D images of platelets by applying direct stochastic optical reconstruction microscopy (dSTORM). The method demonstrated high signal-to-noise ratios, low cellular auto-fluorescence, and reliable blinking behavior. We revealed that large platelets are on average 61% larger in volume than small platelets, and the clustering-based curvature parameters of small platelets differ by 70% from those of large platelets. Moreover, within cluster dimensions up to 300 nm, small and large platelets exhibited a similar probability of 0.246, which increased to 0.718 when extending the range to 1000 nm.
Date of Award | 2023 |
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Original language | English |
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Supervisor | Jaroslaw Jacak (Supervisor) |
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Activation study of small and large platelets determined by dSTORM
Milić, S. (Author). 2023
Student thesis: Master's Thesis