Platelet-dependent thrombin generation is a helpful tool to assess ex vivo the interaction between platelets and plasma coagulation factors in the initiation, amplification, and inhibition of thrombin generation (TG). This review article discusses the most relevant available data on the clinical applications of fluorogenic TG, the most widely used TG assay, performed in the presence of platelets, i.e., in platelet-rich plasma. With respect to prothrombotic states, arterial hypertension and obesity were the most prominent cardiovascular conditions linked to increased platelet-dependent TG. In addition, platelet-associated hypercoagulability, assessed by the TG assay, has been shown in individuals with active cancer. In terms of bleeding, platelet-dependent TG has been applied to assess bleeding risk in individuals with hemophilia, von Willebrand disease, and Glanzmann thrombasthenia as well as in subjects with other congenital or acquired coagulation factor deficiencies. In addition to risk prediction, a role of the TG assay has been suggested in monitoring antiplatelet therapy in prothrombotic conditions and replacement therapy in bleeding diathesis. Finally, for the routine clinical use and as a biomarker of disease development and progression, better standardization and clinical validation of platelet-dependent TG are still needed.readings from a fluorometer are recorded and calculated by means of the Thrombinoscope software (Thrombinoscope BV). In principle, the measurements in PRP are assessed in fresh material, whereas TG in PFP is assessed in frozen, stored material. For TG measurements in PRP, coagulation is triggered by adding 20 µL exogenous tissue factor (TF) to 80 µL PRP (with adjusted platelet concentration of 150,000 platelets/µL), whereas for TG measurements in PFP, 20 µL exogenous TF and 4 µM phospholipids are added to 80 µL plasma. After prewarming for 10 min at 37 • C in the fluorimeter, the reaction is started by adding 20 µL of a low-affinity fluorogenic substrate for thrombin (Z-Gly-Gly-Arg-AMC) and calcium chloride mixture (FluCa). TG is measured as a function of a calibration signal obtained in a sample from the same plasma (PRP or PFP) after addition of a fixed amount of thrombin-α2-macroglobulin complex (20 µL of thrombin calibrator) and 20 µL of FluCa. The TG method is extensively reviewed in [5][6][7]. Parameters derived from a TG curve and most frequently reported in the studies are lag time, time to minimum thrombin formed expressed in minutes (min); peak height, maximum concentration of thrombin formed expressed in nM thrombin; and endogenous thrombin potential (ETP) or area under the curve expressed as nM of thrombin formed per minute.The aim of this article was to review the available evidence for the potential clinical application of platelet-dependent TG in both prothrombotic and bleeding conditions, as depicted in Figure 1. Aspects preventing its broader application and routine clinical practice will also be discussed as uncertainties and limitations of thrombin generation in the...