Fig. 1

Vasomotion analyses of cerebral vessels from mice from mice before and after anesthesia. (A) Experimental design of the vasomotion detection in awake and anesthetized mouse. (B) Representative still-frame of arteriole in vivo under two-photon microscopy. The yellow solid line represents the line-scan position. (C) Kymographs of pial arterioles through a cranial window in a head-fixed mouse. (D) Representative time-series amplitude ratio traces of diameter (ΔD/D₀) and radius (ΔR/R₀) in a mouse arteriole. (E) The representative Fourier plot across diameter and radius of awake and anesthetized mouse arterioles revealed a broad range of frequencies (0–11 Hz), with a distinct peak centered at around 0.10 Hz (myogenic vasomotion), 5.7 Hz (hypothetical heartbeat in anesthetized mouse) and 9.9 Hz (hypothetical heartbeat in awake mouse). (F, G) The average Fourier plot across diameter and radius of awake and anesthetized mouse (n = 32 arteriole diameter segments and n = 64 arteriole radius segments in 4 awake mice) revealed a broad range of frequencies (0–13 Hz). Blue band shows myogenic vasomotion (~ 0.10 Hz), no band shows hypothetical respiration in diameter index, gray band shows hypothetical respiration of anesthetized mouse in radius index (~ 0.90 Hz), red band shows hypothetical anesthetized heartbeat (2.80–5.95 Hz) and green band shows awake hypothetical heartbeat band (9.70–12.70 Hz in diameter and radius index). The hypothetical heartbeat frequency bands are more pronounced in radius index. (H) Representative still-frame of venule in vivo under two-photon microscopy. (I) Kymographs of venules under awake and anesthetized states. (J) Representative time-series amplitude ratio traces of diameter (ΔD/D₀) and radius (ΔR/R₀) under awake and anesthetized states. (K) The representative Fourier plot across diameter and radius of venules revealed a broad range of frequencies (0–11 Hz), the diameter index only shows the myogenic vasomotion frequency at around 0.10 Hz, while the radius index shows a distinct peak centered at around 0.10 Hz (myogenic vasomotion), 4.05 Hz (hypothetical heartbeat in anesthetized mouse) and 10.1 Hz (hypothetical heartbeat in awake mouse). (L, M) The average Fourier plot across diameter and radius of awake and anesthetized mouse (n = 8 venule diameter segments and n = 16 venule radius segments in 4 awake mice) revealed a broad range of frequencies (0–14 Hz), the diameter index only displays a small amount of myogenic vasomotion frequency band of awake mice, the hypothetical heartbeat frequency band of anesthetized mice and awake mice, while the radius index shows a distinct peak centered at around 0.10 Hz (myogenic vasomotion), 2.80–3.80 Hz (hypothetical heartbeat in anesthetized mouse) and 8.40–11.95 Hz (hypothetical heartbeat in awake mouse).