Coagulopathic resuscitation model establishment in vitro. (A) Schematics of coagulopathy induction and resuscitation, and subsequent workflow. WB was diluted 3:7 with normal saline to induce dilutional coagulopathy (dWB) or was dosed with 75 ng/mL tPA to induce hyperfibrinolytic coagulopathy (lyWB). Cryo or FibCon was mixed 1:5 with dWB and lyWB, providing resuscitation and yielding r:dWB and r:lyWB, respectively. (B) Microfluidic assay setup schematic. Sample syringe containing resuscitated coagulopathies (rCryo:dWB, rCryo:lyWB, rFibCon:dWB, rFibCon:lyWB) was perfused via syringe pump for constant flow rate perfusion through the microfluidic flow chamber, and chambers were imaged in real time using an inverted fluorescent microscope. Images in panels A and B were created by K.A.T. and S.M.S. using BioRender.com. (C) Computational fluid dynamics simulation yielding flow streamlines through the microfluidic chamber (top). Representative phase images (bottom left) from a WB control at 500 s⁻¹ that resulted in a BT of 4.7 minutes (bottom right). (D) BTs of dWB and lyWB upon perfusion through the microfluidic model of bleeding; assay cutoff of 1200 seconds determined by a quadrupling of the mean of WB controls. Data are reported as mean (standard deviation). N = 2 to 3. PRP, platelet-rich plasma; PPP, platelet-poor plasma; Q, flow; TF, tissue factor; tPA, tissue plasminogen activator.