Fig. 3: Bidirectional, in vivo optogenetic regulation of mice preference or aversion behaviors with wirelessly operated, dual-color microscale LED (micro-LED) probes.

a Diagram illustrating the combined viral injection (AAV2/9-CAG-DIO-ChrimsonR-mCherry and AAV2/9-EF1a-DIO-stGtACR2-EGFP) followed by implanting the micro-LED probe into the VTA of DAT-cre mice. b Micrographs of the VTA showing co-expression of stGtACR2 and ChrimsonR, with a dual-color micro-LED probe embedded into the tissue. c Illustration of the real-time place preference test with a two-compartment arena, where a mouse receives signals for dual-color control and its locomotion is recorded by a camera. d Patterns used for optogenetic modulations, including a 10-min pretest, a 30-min red LED stimulation (20 Hz, 10-ms pulse, current 7 mA), a 30-min rest, and a 30-min blue LED stimulation (continuous, current 5 mA). e Representative heatmaps showing real-time preference and aversion behavior following red or blue stimulation for mice co-expressing stGtACR2 + ChrimsonR (experiment, or Exp) or EGFP + mCherry (control, or Ctrl). f Summary of preference indices (the ratio of the time that mice spend in the left chamber to the whole recorded time) under red and blue stimulations for Exp and Ctrl groups (n = 4 mice each). Two-way ANOVA, Sidak’s multiple-comparisons test (baseline versus stim), *P < 0.05, **P < 0.01, ***P < 0.001, n.s. P > 0.05. g Preference indices measured at different times under red and blue stimulations for Exp and Ctrl groups (n = 4 mice each). h Preference indices measured at different times under red and blue stimulations for groups expressing only ChrimsonR or stGtACR2 (n = 3 mice each). All data are represented as mean ± s.e.m. See Supplementary Table 1 for detailed statistical analysis. Source data are provided as a Source Data file.