Centrifugal microfluidic system for functional cellular analysis

Florida State University, Tallahassee FL

Investigators

Abstract

Spheroids and organoids are self-assembled, multicellular, 3D structures that better mimic in vivo tissues than 2D cell cultures. They have become an indispensable in vitro tool for processes such as drug assays, disease modeling, and for mechanistic studies. The secretion of specific factors by spheroids and organoids is vital for accurate recapitulation of in vivo function. Methods to measure secretion from these in vitro systems are indirect, slow, laborious, and combine many spheroids / organoids together masking the dynamics of secretion from the individuals. In this proposal, we will develop technology that will enable online measurement of secretion dynamics from many individual spheroids or organoids in parallel. To accomplish this work, we will perform the following specific aims. In aim 1, a centrifugal system will be developed to deliver a continuous flow of perfusion solution to spheroids / organoids as well as immunoassay assay reagents specific for the secreted factor of interest. These solutions will be delivered via centrifugal pumping to a mixing channel where fluorescence anisotropy will be used to quantify the secreted factor as the device spins. The spinning system and fluorescence anisotropy system will be fully optimized. In aim 2, this system will be applied to several model spheroid systems for validation of the technology. The system will enable quantitative measurement of secretory function multiple times per second from each organoid, making a significant and vertical advance compared to current technology. We anticipate the new technology will be a powerful tool in the field of spheroid and organoid research.