Extended Data Fig. 8: Transient angiogenesis and vascular dynamics in awake and anesthetized states.
From: Long-term optical imaging of the spinal cord in awake behaving mice
a, Individual frames across imaging sessions show onset and reversal of angiogenesis in the spinal cord of a CX3CR1-EGFP mouse. Scale bar, 300 µm. b, Change in spinal cord vessel diameter between general anesthesia and awake states in a CX3CR1-EGFP mouse. Middle row illustrates the same frames after application of a Hessian-based Frangi vesselness filter that highlights the dorsal vein and a subset of dorsal ascending venules. These filtered images are used to calculate changes in vessel diameter. Scale bar, 300 µm. c, Procedure for determining diameter of dorsal vein and ascending venules: a Frangi filter was applied to highlight vessels and their local thickness was then calculated to determine vessel diameter. Example frames are illustrated across three major behavioral states of a Thy1-GFP mouse during a 25-min imaging session. Scale bar, 300 µm. d, Temporal change of vessel diameter and whole-frame fluorescence (normalized to 4-min awake baseline) within a single imaging session in a Thy1-GFP mouse before and after induction of general anesthesia (2% isoflurane). Same as Fig. 3p, but here additional right and left dorsal ascending venules are shown. e, Correlation of dorsal vein diameter and fluorescence during a 25-min imaging session across several behavioral states: awake (red), induction and maintenance of general anesthesia (green, isoflurane 2%), and waking up (emergence) from general anesthesia (blue). First order polynomial best-fit lines and R2 indicated by darker colored lines and associated text, respectively.
