High-Throughput Functional Characterization of Visceral Afferents by Optical Recordings From Thoracolumbar and Lumbosacral Dorsal Root Ganglia

Functional understanding of visceral afferents is important for developing the new treatment to hypersensitivity and pain. The sparse distribution in dorsal root ganglia (DRGs) has challenged conventional electrophysiological recordings. Alternatively, Ca 2+ indicators like GCaMP6f allow functional characterization by optical Here we report a turnkey microscopy system that enables simultaneous imaging at two parallel focal planes from intact DRG. By using consumer-grade components, cost-effective can be made broadly available without loss capacity. It records low-intensity fluorescent signals wide field view (1.9 × 1.3 mm) cover whole mouse DRG, with high pixel resolution 0.7 micron/pixel, fast frame rate 50 frames/sec, capability remote focusing perturbing sample. scanning range (100 motorized sample stage allows convenient recordings multiple DRGs thoracic, lumbar, sacral vertebrae. As demonstration, characterized mechanical neural encoding innervating distal colon rectum (colorectum) mice driven VGLUT2 promotor. A post-processing routine developed conducting unsupervised detection afferent responses recordings, which also compensates motion artifacts caused stimulation colorectum. reported offers solution high-throughput activities large volume DRG tissues. We anticipate application this expedite our innervations.