Figure 7: Schematic illustration of OSCAR workflow.

The brain slice cultures were prepared in organotypic fashion obtained from neonatal mice and embedded in compressing lip and sliced. The slices were incubated with either normal or prion-infected brain homogenates for 1 h, then washed and cultured with liquid air interphase on membrane inserts. Treatments with compounds of interest began at 14 dpi in this study and slices were harvested at 31 dpi. Slices were homogenized and dilutions were used to seed RT-QuIC reactions. RT-QuIC assay was performed in 96-well plate using recombinant prion protein as a substrate with continuous shake/rest cycles. The seeding activity was monitored in real time based on seeding using a thioflavin T (ThT) fluorescence readout and quantified. Positive wells can be clearly visualized based on the traces that cross the threshold fluorescence and by the rapid increase in intensity. The well-area scan image shows the intense amyloid deposits in the well.