Nusieve gtg

All animal operations complied with the protocols approved by the Institutional Ethical Committee of Animal Experimentation of Shenzhen University. C57 mice were purchased from Guangdong Medical Laboratory Animal Center. Before 2-photon imaging, craniotomies were performed on C57 mice (female, 8 to 10 weeks old, weighing 20 to 21 g) centered at 2 mm lateral and posterior to the bregma point. A cover glass was used to seal the cranial window. To fix the mice, a home-made alloy ring was stuck onto the skull by dental cement. During surgery and imaging, a gaseous anesthesia system (Matrix VIP 3000, Midmark) and a heating blanket were used to anesthetize mice and maintain their body temperature, respectively. A prepared aqueous solution ( $200\mu \mathrm{L}$ ) of fluorescent probes TPETPABT²⁶ (link) was injected through the orbit to label the brain vasculature.
Zebrafish embryos of the Tg(kdrl:EGFP) strain were obtained from the China Zebrafish Resource Center and raised in E3 solution containing 0.003% $N$ -phenylthiourea (Sigma) to inhibit pigmentation after 20 h postfertilization. Vascular epithelial cells were labeled with EGFP. Prior to live imaging, zebrafish were anesthetized with $600\mu \mathrm{M}$ Tricaine (Sigma, Cat. # E10521) and mounted in 1% low-melting-point agarose (NuSieve GTG, Cambrex BioScience, Cat. # 50080) for imaging to minimize movement artifacts during imaging.