Fig. 3
From: Zoledronic acid alters hematopoiesis and generates breast tumor-suppressive bone marrow cells
Effects of a single dose of zoledronic acid (ZOL) on bone marrow vasculature. a-d Bone marrow vasculature was visualized 3 days after control (Ctl) or ZOL treatment using immunofluorescence staining against the vascular endothelial cell marker endomucin (green) on 30-μm-thick sections of gelatin-embedded tibiae: 8 × 8 tile scans captured with × 20 objective using the Nikon Eclipse Ti, NIS-Elements-software Version 4.30, CFI Plan Fluor 20x MI (NA 0.75); scale bars 250 μm, arrows indicate branched vasculature, dashed lines represent growth plate demarcation, Ep epiphysis, Met metaphysis, Dia diaphysis. e-f Z-stacks with a depth of 20 μm of the metaphyseal vasculature (e) and confocal images of the diaphyseal bone marrow vasculature (f), acquired with the Nikon A1 confocal microscope, NIS-Elements-software Version 4.30, CFI Plan Fluor 20x MI (NA 0.75), Endomucin-positive vascular endothelial cells (green; Alexa555), nuclei (blue; 4',6-diamidino-2-phenylindole (DAPI)); scale bars 100 μm; n = 3/group. Phase-contrast image (right) is a toluidine blue stain of a nude mouse tibia to indicate the regions of imaging for the vasculature stains. g Average mouse plasma vascular endothelial growth factor (VEGF) concentration (pg/mL) 3 days after a single dose of ZOL or Ctl treatment (n = 4 for Ctl, n = 3 for ZOL). ns not significant