Morgen Leithe
Research Mentor: Benjamin Singer
Mentor Department: Internal Medicine / Pulmonary and Critical Care Medicine, Medicine
Author(s): Morgen Leithe, Andrew Stephens, Benjamin Singer
Session: Session 7 (4:00 PM – 4:50 PM)
Presentation Type: Oral Presentation
Abstract
This project focuses on the development of a microfluidic chip for performing multiplex ELISA with small (<5?µL) sample volumes. Traditional immunoassays, such as ELISA, often lack the sensitivity needed to detect low-abundance protein biomarkers in diluted samples. Digital immunoassays, which leverage single-molecule quantification, provide improved sensitivity and can be integrated into microfluidic systems, enabling multiplexing. The Sepsis Survival Lab is investigating the use of these immunoassay technologies for serial, repeated measurements of blood cytokine concentrations in murine sepsis models where sample volume is limited. The minimal sample volume requirement allows cytokine measurement via tail vein blood sampling, which can be performed several times during an experiment without sacrificing the animal, unlike conventional cardiac puncture. It is not well understood whether the location of cytokine sample collection in mice affects signal output, making it difficult to discern whether differences are due to cytokine concentration or variability introduced during sampling. To address this, the Sepsis Survival Lab conducted experiments comparing cytokine measurements from tail vein and cardiac puncture in murine sepsis models. Identifying statistically significant differences between sampling sites is necessary for accurate interpretation of cytokine data. The findings suggest there is insufficient evidence to conclude that cytokine concentrations differ significantly between tail vein and cardiac puncture samples, indicating that variability is more likely due to sampling or pipetting errors. In future experiments, personnel can use either tail vein or cardiac sampling with careful attention to sample volume. Next steps will involve characterizing sampling error and investigating other potential confounding variables in human blood samples.
