Platelet-platelet aggregates at single-event resolution as parameter in health monitoring

Platelet analysis is crucial to assess in hemostasis, health, and disease. Current diagnostics primarily analyze bulk platelets, limiting the assessment of subtle, single-aggregate changes. We utilize Plateaggrate, a novel neuronal network-driven tool, for scoring platelet-platelet aggregation at single-event resolution using imaging flow cytometry (IFC) of labeled whole blood. This study evaluated Plateaggrate in 96 healthy human probands (61 males, 35 females; median age 39) to identify age and sex dependencies in native and ADP-stimulated platelets. Antibody-labeled platelets (CD42b and CD62P) were analyzed using IFC and segmented into singlets, doublets, triplets, and multiplets. While the weighted platelet aggregation score (WPA) showed no sex or age dependencies, Plateaggrate identified a slight but significant negative correlation of platelet singlets and total platelets with age. ADP-mediated activation, however, yielded robust platelet activation and aggregation independent of age, sex, and aggregate nature. In summary, this study validates the suitability of the WPA platelet algorithm for single-event platelet aggregation analysis in future disease patient cohorts.