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Applicable species: mice (C57BL/6, FVB, etc.), rats (SD, Long-Evans), NHP (rhesus/cynomolgus) [12-1]. Dose range: adult mouse striatum 1×10^9-5×10^10 vg/site, hippocampus 1×10^9-1×10^10 vg/side; rat striatum 2-6×10^10 vg/site; NHP putamen 1.5×10^11 vg/brain (2×50 μL) [12-1]. Injection volume: mouse 0.2-2 μL/site (cumulative ≤5 μL/brain); rat 0.5-2 μL; NHP 50-150 μL/site[12-1]. Needle: Mouse Hamilton 7000 series equipped with 33 G stainless steel needle or glass microelectrode; NHP customized fused-silica anti-reflux catheter [12-1]. Speed: 100-200 nL/min is most commonly used for mice; NHP ramp-up 0.2→0.5→1.0→3.0 μL/min[12-1,12-40]. Anesthesia: Mice were treated with isoflurane 3-5% for induction and 1-2% maintenance, intraoperative thermostat and postoperative buprenorphine analgesia; NHP isoflurane 1-3% for tracheal intubation monitoring [12-1]. Key points of operation: The height difference between bregma and lambda is <0.05 mm, drill and thin the skull to open the dura mater, leave the needle for 5-10 minutes after injection and then withdraw the needle at 1 mm/min to reduce reflux [12-1]. Coverage: Diffusion radius is about 1-2 mm (half-life distance 500-1000 μm), limited to around the injection site + anterograde/retrograde projection pathway; cell type is determined by the promoter; peak expression is 2-4 weeks [12-1]. Safety: needle tract bleeding, high-dose epilepsy in cortex/hippocampus; direct neurotoxicity has been reported in NHP intracerebral injection of AAVrh8 [12-38].