Adhesion and spreading of ΔneoΔHS platelets on collagen under static and flow conditions. (Ai) Washed murine platelets (1 × 107/mL) from wild-type (top row) and ΔneoΔHS (bottom row) mice were allowed to adhere and spread on immobilized fibrinogen (Fibrinogen) and collagen (Collagen). Adherent platelets were fixed, permeabilized, and stained with rhodamine-phalloidin. Platelets were also plated on BSA-covered coverslips (BSA). Adherent platelets were imaged using fluorescence microscopy, and cropped representative images are shown. Images taken are from 1 experiment, representative of 3. (ii) Graphs show the number of platelets adherent to the thrombogenic matrix (left) and the number of platelets that had undergone some change in shape, recorded as a percentage of spread platelets (right), mean ± SEM. ▪ indicates WT platelets; □, ΔneoΔHS platelets. (B) Whole blood was obtained from individual wild-type (Wild type) or ΔneoΔHS (ΔneoΔHS) mice and perfused over collagen-coated (100 μg/mL) capillary tubes at a shear rate of 800 s^-1 for 2 minutes followed by Tyrode buffer for 5 minutes. Five fields were imaged (× 63 objective), and representative images of adherent platelets and aggregates from 2 individual wild-type and ΔneoΔHS mice are shown. These data are from 1 flow experiment representative of 2. On the right, red blood cells, PRP and PPP were isolated from wild-type whole blood. Blood was reconstituted to represent either 100% (Wild type reconstituted, 100%) or 20% (Wild type reconstituted, 20%) of the original wild-type platelet count in whole blood. These reconstituted samples were perfused and treated as whole blood flows, described in earlier part of (B). Note that the white regions of the images correspond to regions of substantial and high aggregates. The increased height has induced flare in the phase-contrast image.