ORGAN-ON-A-CHIP PLATFORM WHICH ENABLES SUPER RESOLUTION MICROSCOPY

Parkinson’s disease (PD) is a neurodegenerative disease known for abnormal accumulation of aggregated misfolded α-Synuclein (α-Syn) in the central nervous system (CNS). While these aggregates are a hallmark of the disease, many fundamental questions, which relate to the formation and propagation dynamics of these aggregates, remain unknown. One of the major challenges is the ability to have both spatial information in high resolution, and the ability to monitor the α-Syn propagation between multiple cell types.

To overcome the above challenge, we developed a modular Organ-on-a-Chip platform that enables us to conduct super-resolution microscopy (SRM). This platform allows us to co-culture multiple cells type and identify propagation of α-Syn between two separated cell populations, and monitor the system with functional tools (such as a multi-electrode array) and SRM.

The new platform tackles several fundamental challenges in characterizing neuronal cells in vitro. The system allows us to co-culture cells on two sides of a membrane to assess the cell-cell communication, and simultaneously to assess their functionality. In addition, it overcomes the challenge of SRM imaging, which is mostly limited by the short working distance (170um), by the ability to take out the chip membrane while keeping the integrity of the tissue.

In this work, we present a case study, where we co-culture neurons which overexpress α-Syn together with naïve neurons and we monitor via direct Stochastic Optical Reconstruction Microscopy (dSTORM) the dynamics of the aggregation and propagation of α-Syn from the α-Syn expressing neurons to the naïve ones. We applied a modified cluster analysis algorithm (DBSCAN) on the dSTORM images to characterize the precise size, shape, density, and composition of α-Syn aggregates in both neuronal populations.