title: Measuring angiogenesis and hemodynamics in mice.
abstract: This protocol outlines methods to measure vascular parameters, including angiogenesis (e.g., vascular density, length, diameter) and hemodynamics (e.g., erythrocyte velocity), in tumors and normal vascular networks in mice. Fluorescein-isothiocyanate (FITC)-dextran is injected into the tail vein of mice to visualize microvessels within 150 μm (using single-photon microscopy) or ~600 μm (using multiphoton laser-scanning microscopy [MPLSM]) of a tumor/window interface. Randomly selected areas (three to six locations/tumor or animal) are investigated using long-working-distance objectives with appropriate magnification. During each observation period, FITC-fluorescence images are recorded for 60 sec, and the videotapes are analyzed off-line for single-photon microscopy; or a three-dimensional (3D) image stack of the vessel network is generated, and vessel properties are measured for MPLSM. If desired, red blood cell (RBC) flux can be measured on a vessel-by-vessel basis using fluorescent tracer RBCs.
authors.forename: Rakesh K
authors.initials: RK
authors.lastname: Jain
authors.forename: Lance L
authors.initials: LL
authors.lastname: Munn
authors.forename: Dai
authors.initials: D
authors.lastname: Fukumura
mesh_terms: D061605:Administration, Intravenous; D000818:Animals; D003911:Dextrans / Q000008:administration & dosage; D004195:Disease Models, Animal; D016650:Fluorescein-5-isothiocyanate / Q000008:administration & dosage / Q000031:analogs & derivatives; D006439:Hemodynamics / Q000502:physiology*; D007091:Image Processing, Computer-Assisted / Q000379:methods; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods; D018715:Microscopy, Video / Q000379:methods; D009369:Neoplasms / Q000473:pathology*; D009389:Neovascularization, Pathologic; D018919:Neovascularization, Physiologic; D013194:Staining and Labeling / Q000379:methods
chemical_list: D003911:Dextrans; C015219:fluorescein isothiocyanate dextran; D016650:Fluorescein-5-isothiocyanate