ABSTRACT Premature infants are frequently at risk of infections, jaundice and hypoglycemia – conditions monitored by analysis of blood for c-reactive protein (CRP), bilirubin and glucose respectively. Typically, 0.5 mL of blood is drawn each time an infant is tested, with multiple blood draws happening over the course of days. Given that a total blood volume for premature infants may be as little as 50 mL, up to 20% of blood volume may be lost due to testing. This puts infants at risk of iatrogenic anemia and packed red blood cell transfusions. While these transfusions are considered safe in many regards, they do expose neonates to transfusion-acquired infections, cytomegalovirus in particular. Our team has identified a need for a volume-sparing blood analysis platform that could be deployed in neonatal intensive care units (NICUs). During Phase I of this STTR application, we de- veloped a valve-enabled microfluidic device capable of isolating plasma and analyzing multiple biomarkers, including glucose, from ~5 microliters of blood in under 10 min. Our device used ~100 fold less blood and de- livered results 10 times faster compared to current assays while eliminating the need for benchtop laboratory equipment (e.g. centrifuge for plasma separation). The microfluidic devices were prototyped in silicone rubber (PDMS). During Phase II of the project, we will fabricate automated microfluidic devices using materials ame- nable to scalable and inexpensive manufacturing. Our devices will be capable of detecting CRP, bilirubin and glucose and will be suitable for operation at the point of care, in the NICU. Broader impacts: The microfluidic device and control system to be developed in this project represent a plat- form technology with broad utility for monitoring a wide range of analytes from a small blood volume. This means that, in the future, the multiplexing capabilities of this technology may be enhanced to incorporate other analytes relevant to neonatal health including but not limited to inflammatory cytokines for detection of sepsis, potassium ions, calcium ions, and BUN.