PROJECT SUMMARY Blood coagulation is a complex process that involves a balance of plasma proteins and platelets. A slight imbalance in any one of these can cause life-threatening bleeding or clotting. Current treatments for bleeding disorders such as hemophilia include recombinant or plasma-derived factor replacement therapies, factor mimetic therapies, and bypassing agents, but they can cause serious side effects and cost in excess of $1,000,000 per patient per year. Effectively dosing a treatment for hemophilia is often problematic due to the narrow therapeutic window between bleeding and coagulation, numerous food and drug interactions, and high variability in dose response in different people. Patients require frequent monitoring of the coagulation status, however, there is no single test currently available to provide comprehensive information about coagulation in a convenient way, and existing tests are not as helpful for modern hemophilia treatments as they are for previously gold-standard treatments. Our revolutionary acoustic tweezing platform has the capability to simultaneously measure early, intermediate, and late stages of coagulation and requires only a single drop of whole blood per test. The platform utilizes acoustic levitation to manipulate a tiny drop of a sample and measure the change in properties while the blood drop coagulates. This technology has the potential to replace many existing coagulation tests and has a form appropriate for use in outpatient clinics, making testing less invasive and more convenient for patients. This technology also addresses the current unmet in the market for coagulation testing in neonatal, pediatric, and other vulnerable patient populations, who suffer from serious side-effects related to excessive and repetitive blood draws. Additionally, the data output by the acoustic tweezing platform is comprehensive, allowing for more capable monitoring of drugs to reduce the costs of treatment and prevent adverse drug effects. The objective of this SBIR Phase I project is to develop a point-of-care coagulation analyzer that can provide coagulation profile of patients with hemophilia using a single drop of whole blood. The new prototype will be designed to perform experiments via acoustic tweezing spectroscopy (ATS), which currently require a large, lab-based prototype. The two specific aims proposed in this application will cover 1) development of the benchtop prototype from current lab-grade electronics and 2) evaluation of the device on blood samples collected from healthy subjects and patients with hemophilia.