Imaging Striatal Dopamine Release Using a Non-Genetically Encoded Near-Infrared Fluorescent Catecholamine Nanosensor

Abraham G. Beyene; Kristen Delevich; Jackson Travis Del Bonis-O’Donnell; David J. Piekarski; Wan Chen Lin; A. Wren Thomas; Sarah J. Yang; Polina Kosillo; Darwin Yang; Linda Wilbrecht; Markita P. Landry

Raw movie files were processed using a custom-built MATLAB program (available for download: https://github.com/jtdbod/Nanosensor-Brain-Imaging). Briefly, for each raw movie stack (600 frames), a per pixel (F-F0)/F0 was calculated using the average intensity for the first 5% of frames as F0, and F represents the dynamic fluorescence intensity at each pixel. Regions of interest (ROIs) were identified by calculating a median filter convolution and then performing thresholding using Otsu's method followed by a morphological dilation operation. ∆F/F traces were then calculated for each generated ROI. ROI sizes were computed using the measured pixel area and approximating each as a circle to calculate an equivalent radius.