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Over the past fifteen years my lab has used microfluidic technologies to re-create 3D microenvironments that mimic human tissues (microphysiological systems or “organ-on-a-chip” technology) and to isolate rare and specific immune cells in the peripheral circulation. The former has important applications in drug development, toxicity screening, and cell/extracellular vesicle trafficking in tumor metastasis, while the latter can provide insights into the immune response to viral challenges, vaccines, and cancer. We began by creating 3D perfusable microvascular networks, which ultimately led to the creation of an active start-up company, Aracari Biosciences, based in Irvine, CA. From there, our focus has largely been on the creation of 3D vascularized microenvironments of cancer, bone marrow, and the extracellular matrix to enhance our understanding of immune cell and extracellular vesicle trafficking. More recently, we have utilized microfluidic devices to capture rare and specific lymphocytes (both B cells and T cells) to address compelling questions related to immunity, in particular the response of memory B cells to a viral challenge and cytotoxic CD8+ T cells to a tumor. This seminar will provide an overview of our core technologies and some of the key findings related to immune cell and extracellular trafficking in 3D tissue microenvironments.