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Hemostasis depends on the coordinated interaction between platelets, coagulation factors, endothelium and shear forces. Current point-of-care (POC) assays evaluate isolated components of haemostasis or operate under nearly static conditions, limiting their ability to reproduce physiological thrombus formation. In this study, we performed the technical validation of Smart Clot, a fully automated, microfluidic POC platform that quantifies platelet aggregation, fibrin formation, and total thrombus growth under controlled arterial shear using unmodified whole blood. Recalcified citrated blood was perfused over collagen at \( \dot{\gamma} \)w = 300 s⁻¹. Dual-channel epifluorescence microscopy acquired platelet and fibrin(ogen) signals at 1 frame per second. Integrated Density time-series were fitted with a five-parameter logistic model; first derivatives and their integrals yielded standardized pseudo-volumes for platelets, fibrin, and total thrombus. Sixty-two healthy donors established reference distributions; one-hundred-thirteen patients on antithrombotic therapy assessed pharmacodynamic sensitivity. Platelets-derived parameters were approximately normally distributed, whereas fibrin(ogen) and total-thrombus values followed log-normal patterns. Anticoagulants and antiplatelet agents produced graded, mechanism-consistent inhibition of all thrombus components. Cardiopulmonary-bypass samples showed profound but transient suppression of platelet and fibrin activity. Across activity ranges, multiple statistical assessments supported high analytical repeatability. Smart Clot provides rapid, reproducible, flow-aware quantification of platelet–fibrin dynamics, capturing pharmacological modulation and peri-operative impairment with high sensitivity. These results support its potential as a next-generation POC assay for physiological hemostasis assessment.