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Steady-State Blood Volume Measurements in Experimental Tumors with Different Angiogenic Burdens—A Study in Mice¹

¹ From the Center for Molecular Imaging Research, Massachusetts General Hospital and Harvard Medical School, Bldg 149, 13th St, 5406, Charlestown, MA 02129. Received January 9, 2002; revision requested February 4; revision received April 2; accepted May 23. Address correspondence to R.W. (e-mail: weissleder@helix.mgh.harvard.edu).

View larger version (60K): [in a new window]   Figure 1. Intravital microscopic images enhanced with fluorescently labeled monocrystalline iron oxide 46 depict 9L tumor neovasculature. The first two images from the left are, respectively, low- (green fluorescent protein; magnification, x2) and high- (green fluorescent protein; magnification, x60) resolution images that show the vessels dark in contrast to the fluorescent tumor cells expressing green fluorescent protein. The contrast is accentuated because hemoglobin in blood vessels is an efficient absorber of fluorescent light. The third to sixth images from the left represent a time series of near-infrared fluorescent (NIRF) images obtained in the indocyanine 5.5 channel, in which the fluorescently labeled LCDIO within vessels appears bright. Note that the distribution of the intravascular probe (ie, indocyanine 5.5-labeled monocrystalline iron oxide 46) is constant during the observation time of 30 minutes; this indicates that there is no leakage. (Indocyanine 5.5-labeled monocrystalline iron oxide 46; magnification, x60.)

View larger version (164K): [in a new window]   Figure 2. CD31 immunohistochemical analysis of the four experimental tumor models. Hotbeds of tumoral microvessels are stained in blue (CD31) against a red tumoral background. The microvessels in the different tumor models have different densities and morphologic appearances. (Nuclear fast red counterstain; magnification, x20.)

View larger version (34K): [in a new window]   Figure 3. MR images of the four experimental tumor models. Top row: Color-coded VVF tumoral vascularity maps are superimposed onto the tumors and derived from pre- and postcontrast T2*-weighted MR imaging sequences. VVF values are from 0% to 30%. Note the heterogeneity of VVF among the tumor models. Bottom row: The different tumors are outlined on postcontrast T1-weighted spin-echo MR images (300/6).

View larger version (24K): [in a new window]   Figure 4. Graphs illustrate heterogeneity among image section-selected VVFs of different tumor models. For each model, left and right tumors within the same animal are plotted along image sections of the entire tumor. Note the considerable heterogeneity among measurements between the left and right tumors and among the different image sections of ipsilateral tumors. These changes were most dramatic in the EOMA tumors.