We next compared the frequency gating of the forward scatter data of the PLT baseline control condition (t0) with the PLTs incubated with IONFs-C1 for 30 minutes, 3 hours and 6 hours (Figure -C and Figure ). It is notable that in case of the 30-minute incubation the trend of the frequency ratios, gated on the histogram plots of forward scattering, is opposite to that of baseline condition. Specifically, the fr2 frequency gate is smaller than the fr3 frequency gate for 30-minute incubation, whereas fr2 is larger than fr3 for the baseline condition. However, for longer incubation times, the frequency trend is similar to that of the baseline, confirming that PLT activation is limited in time and that exposure to IONFs-C1 does not induce PLT apoptosis. ). For example, for 30 minutes, in the same ROI, the addition of the IONFs to RBCs and PLTs induced a decrease in T1 and T2 relaxation times. Indeed, T1 relaxation times of the solutions of RBCs (691.60 ± 6.56 ms) and PLTs (675.76 ± 5.76 ms) with IONFs-C1 (Figure -A and Table In agreement with previous studies, it was possible to identify in both the control and IONFs-C3 an abundance of glycogen granules (spheroidal formations about 50 nm in size) contained within the PLT cytoplasm. glycogenolysis. In the case of IONFs-C1, individual or aggregated IONFs were seen both in the extracellular and intracellular environment, as indicated by brown arrows. This suggests that the IONFs-C1 has induced transition to a high-demanding state where the stored glycogen is engaged. ) Furthermore, we concluded that IONFs-C1 at 30 minutes were possibly internalized by endocytosis.