Developing a Midbrain Organoid-on-a-Chip for Live Cell Imaging and Parkinson’s Disease Modeling Team 13: Eugenia Angelopoulos, Victoria Lara-Aguilar, Maria A. Parada R. Advisors: Emma Bortz, Xue Han, Drew Martin Parkinson’s disease (PD) is the second most common neurodegenerative disorder, currently af- fecting an estimated 1 million individuals in the U.S. and more than 10 million people worldwide. PD involves the degeneration of dopaminergic neurons in the midbrain substantia nigra pars compacta (SNc) and is characterized by symptoms like bradykinesia and tremors. The majority of PD cases remain idiopathic but exposure to environmental toxins can lead to Parkinsonism with pathologic hallmarks such as disruption of the lysosome-autophagy system, mitochondrial dysfunction and eventual death of dopaminergic neurons. Researchers have developed human midbrain organoids (MBOs), which are three dimensional in vitro models that hold great potential for investigating the complex cellular variability of the human midbrain and are preferable to other animal models or 2D cell lines. The use of MBOs derived from human neuroepithelial stem cells (hNESCs), allows for faster maturation of MBOs and higher yield of differentiated neural cells. However, no sufficient progress has been made in developing a way to conduct multiple imaging sessions and improve the imaging quality of developing MBOs, while keeping them alive. In this project, we developed four midbrain organoid-on-a-chip (MOC) designs, and tested the most suitable designs-one in- verted and one upright-using MBOs derived from hNESCs to make the organoids stationary and compatible with fixed and dynamic fluorescence imaging using upright microscopy. MOCs may ultimately improve imaging quality and ease, and thus may reveal more information about midbrain cellular structure and dynamics, bringing researchers one step closer to finding a cure for PD..