Fig. 1

Imaging of functional BBB in 3dpf and 6dpf zebrafish. (A,B) The Dextran Texas Red (70,000 MW, 1 mg/ml), Evans Blue Dye (EBD, 961 MW, 5 mg/ml), Lucifer Yellow (LY, 457 MW, 4 mg/ml) and DAPI (350 MW, 1.5 mg/ml) were mixed and injected into the circulation of 3dpf flk:eGFP or flk:mCherry embryos (5 nl/fish, green indicating endothelial cells) from caudal vein (CV). Z-stack confocal living images of the cerebral capillaries and segmental capillaries (Fig. S2A) were obtained at 30–60 min after injection. (A, area1) Dextran and DAPI were restrained in cerebral capillaries (CCTA), but were leaked out from primary head Sinus (A, area2) and segmental capillaries (Fig. S2A). (B) CBD (white) and LY (red, pseudocolor) were restrained in CCTA (mCherry was pseudocolored as green). Areas in dotted boxes were magnified right. (C) DAPI was injected into the circulation of 6dpf embryos (maintained at 28 °C) from CV. Z-stack confocal living images were taken at 1 h post injection. White arrows indicating nucleus of host cells neighboring to vessels; Red arrows indicating endothelial nucleus. (D)DAPI was injected into the circulation of 6dpf embryos (normally maintained 3dpf zebrafish were shifted to 33 °C for 3days) from CV. Z-stack confocal living images were taken at 1 h post injection. White arrows indicating nucleus of host cells neighboring to vessels; Red arrows indicating endothelial nucleus.