bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 24, 2025

Fluorescent sensors have revolutionized the measurement of molecules in brain, and dLight dopamine sensor has been used extensively to examine reward- cue-evoked release, but only recently field turned its attention spontaneous release events. Analysis events typically requires evaluation hundreds over minutes hours, most common method analysis, z-scoring, was not designed for this purpose. Here, we compare accuracy reliability three different analysis methods identify pharmacologically induced changes uptake freely moving C57BL/6J mice. The D1-like receptor antagonist SCH23390 prevent from interacting with extracellular space, while cocaine inhibit raclopride increase nucleus accumbens. We examined peak-to-peak frequency, peak amplitude, width, time spent above an established cutoff. were 1) widely-used "Z-Score Method", which automatically smooths baseline drift normalizes recordings using signal-to-noise ratios, 2) a "Manual local baselines adjusted manually individual cutoffs determined each subject, 3) "Prominence Method" that combines z-scoring prominence assessment tag peaks, then returns preprocessed data kinetic analysis. First, drastically reduced number signals detected as expected, when Manual Method used. Z-scoring failed any changes, due amplification noise diminished. Cocaine increased signal width expected Prominence Methods, Z-Score Method. Finally, raclopride- increases amplitude correctly identified by Methods. kinetics. Thus, %ΔF/F values, ideally Method, use z- scoring is appropriate.

Language: Английский