By Addison Norden

Faculty Mentor: Dr. Randall Reif

Abstract

Apoptosis is the tightly regulated process of programmed cell death that eliminates excess, damaged, or infected cells from the body. It plays a critical role in tissue homeostasis, and mutations affecting this process can result in tumor development, where the damaged cell is unable to trigger apoptosis and continues to proliferate unregulated. Thus, apoptotic triggers are being investigated as potential cancer treatments. To develop these therapies, background information is required on the temporal dynamics of apoptosis, as examined in Reif et al.(1) This research uses microfluidic devices to capture Jurkat T lymphocytes, keeping the cells stationary for the nine-hour experiment. A Y-junction is attached to the microfluidic device channel to alternate providing nutrients and a fluorogenic caspase probe, which allows apoptotic activity to be monitored. However, this research is difficult to continue due to a high instance of experimental failure from air bubbles. The original Y-junction frequently introduced air bubbles to the channel, which removed the captured cells and invalidated all experimental data. A design using needles to introduce solutions to the channel was proposed but is not safe to be used with Jurkat T lymphocytes or other reagents. In this work, I developed a novel Y-junction design utilizing Luer lock connections and catheter-tubing junctions. This successfully eliminated the introduction of air bubbles to the channel, resulting in decreased experimental failure and allowing for research to continue with more reliable experimental results.

(1)Reif, R. D.; Aguas, C.; Martinez, M. M.; Pappas, D. Temporal Dynamics of Receptor-Induced Apoptosis in an Affinity Microdevice. Anal Bioanal Chem 2010, 397 (8), 3387–3396. https://doi.org/10.1007/s00216-010-3567-1.