Semiautomatic Landmark-Based Two-Dimensional—Three-Dimensional Image Fusion in Living Mice: Correlation of Near-Infrared Fluorescence Imaging of Cy5.5-Labeled Antibodies with Flat-Panel Volume Computed Tomography

Connecting fluorescence signals with anatomic structures enhances our ability to monitor biologic processes in mice. Here, we present a semiautomated approach to correlate two-dimensional (2D) noninvasive near-infrared fluorescence (NIRF) imaging with three-dimensional (3D), high-resolution, flat-panel volume computed tomography (fpVCT). We developed an algorithm to colocalize fluorescence signals of NIRF-labeled antibodies directed against matriptase and urokinase plasminogen activator receptor (uPAR) to orthotopic carcinomas in mice visualized by fpVCT. For this purpose, mice were anesthetized and fixed on a multimodality animal bed containing fiducial markers filled with iodine-containing contrast agent and fluorescent dye. After intravenous administration of contrast agent and Cy5.5-labeled antibodies, NIRF and fpVCT images were obtained, without repositioning the mice. Binding of Cy5.5-labeled matriptase-specific antibody to pancreatic tumors and Cy5.5-labeled uPAR-specific antibody to mammary carcinomas was assessed by time-domain NIRF imaging measuring the location of fluorescence intensity and its lifetime. In summary, we developed a novel 2D-3D registration technique for image fusion with NIRF imaging and fpVCT to provide complementary information in tumor models on the in vivo association of functional information with anatomic structures. The combination of fpVCT with NIRF imaging will now allow targeted and effective monitoring of preclinical tumor therapies.

[1]  Jürgen Weese,et al.  A comparison of similarity measures for use in 2-D-3-D medical image registration , 1998, IEEE Transactions on Medical Imaging.

[2]  R Richards-Kortum,et al.  Optical Systems for in Vivo Molecular Imaging of Cancer , 2003, Technology in cancer research & treatment.

[3]  Vasilis Ntziachristos,et al.  Looking and listening to light: the evolution of whole-body photonic imaging , 2005, Nature Biotechnology.

[4]  Eckhardt Grabbe,et al.  Semi-Automatic Classification of Skeletal Morphology in Genetically Altered Mice Using Flat-Panel Volume Computed Tomography , 2007, PLoS genetics.

[5]  Chun Xing Li,et al.  Near-infrared optical imaging of integrin alphavbeta3 in human tumor xenografts. , 2004, Molecular imaging.

[6]  J. Frangioni.  In vivo near-infrared fluorescence imaging. , 2003, Current opinion in chemical biology.

[7]  Ralph Weissleder,et al.  Near-infrared fluorescent nanoparticles as combined MR/optical imaging probes. , 2002, Bioconjugate chemistry.

[8]  Meng Yang,et al.  Real-time optical imaging of primary tumor growth and multiple metastatic events in a pancreatic cancer orthotopic model. , 2002, Cancer research.

[9]  Marta Zientkowska,et al.  Morphologic changes of mammary carcinomas in mice over time as monitored by flat-panel detector volume computed tomography. , 2008, Neoplasia.

[10]  D. Boturyn,et al.  Noninvasive Optical Imaging of Ovarian Metastases Using Cy5-labeled RAFT-c(-RGDfK-)4 , 2006, Molecular imaging.

[11]  J. Kaspareit,et al.  An Orthotopic Model of Ductal Adenocarcinoma of the Pancreas in Severe Combined Immunodeficient Mice Representing All Steps of the Metastatic Cascade , 2001, Pancreas.

[12]  B. Rice,et al.  Quantitative comparison of the sensitivity of detection of fluorescent and bioluminescent reporters in animal models. , 2004, Molecular imaging.

[13]  E. Haber,et al.  Intensity Gradient Based Registration and Fusion of Multi-modal Images , 2007, Methods of Information in Medicine.

[14]  Wolfhard Semmler,et al.  Volumetric computed tomography (VCT): a new technology for noninvasive, high-resolution monitoring of tumor angiogenesis , 2004, Nature Medicine.

[15]  Vasilis Ntziachristos,et al.  Volumetric tomography of fluorescent proteins through small animals in vivo. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[16]  E. Sevick-Muraca,et al.  Near-Infrared Fluorescence Optical Imaging and Tomography , 2004, Disease markers.

[17]  Fred L. Bookstein,et al.  Principal Warps: Thin-Plate Splines and the Decomposition of Deformations , 1989, IEEE Trans. Pattern Anal. Mach. Intell..

[18]  Jennifer L. Harris,et al.  Cellular Localization of Membrane-type Serine Protease 1 and Identification of Protease-activated Receptor-2 and Single-chain Urokinase-type Plasminogen Activator as Substrates* , 2000, The Journal of Biological Chemistry.

[19]  Ronald G Blasberg,et al.  Multimodality in vivo molecular-genetic imaging. , 2004, Bioconjugate chemistry.

[20]  C. Meltzer,et al.  PET/CT: form and function. , 2007, Radiology.

[21]  Wolfhard Semmler,et al.  Potential applications of flat-panel volumetric CT in morphologic and functional small animal imaging. , 2005, Neoplasia.

[22]  K. Uhland.  Matriptase and its putative role in cancer , 2006, Cellular and Molecular Life Sciences CMLS.

[23]  H. Nielsen,et al.  Quantitation of the receptor for urokinase plasminogen activator by enzyme-linked immunosorbent assay. , 1994, Journal of immunological methods.

[24]  W. Cai,et al.  Near-infrared fluorescence imaging of tumor integrin alpha v beta 3 expression with Cy7-labeled RGD multimers. , 2006, Molecular imaging and biology : MIB : the official publication of the Academy of Molecular Imaging.

[25]  Baljit Singh,et al.  Expression of the serine protease matriptase and its inhibitor HAI-1 in epithelial ovarian cancer: correlation with clinical outcome and tumor clinicopathological parameters. , 2002, Clinical cancer research : an official journal of the American Association for Cancer Research.

[26]  R. Weissleder,et al.  Tomographic fluorescence mapping of tumor targets. , 2005, Cancer research.

[27]  Pascal Gallant,et al.  Sensitivity characterization of a time-domain fluorescence imager: eXplore Optix , 2007 .

[28]  W. Bode,et al.  Secondary amides of sulfonylated 3-amidinophenylalanine. New potent and selective inhibitors of matriptase. , 2006, Journal of medicinal chemistry.

[29]  Perry J Pickhardt,et al.  Microcomputed tomography colonography for polyp detection in an in vivo mouse tumor model. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[30]  Eva M. Sevick-Muraca,et al.  Near-Infrared Optical Imaging of Integrin αvβ3 in Human Tumor Xenografts , 2004 .

[31]  Integrated PET/CT in lung cancer imaging: history and technical aspects. , 2007, JBR-BTR : organe de la Societe royale belge de radiologie (SRBR) = orgaan van de Koninklijke Belgische Vereniging voor Radiologie.

[32]  Ralph Weissleder,et al.  In vivo selection of phage for the optical imaging of PC-3 human prostate carcinoma in mice. , 2006, Neoplasia.

[33]  Robert E. Lenkinski,et al.  In vivo near-infrared fluorescence imaging of osteoblastic activity , 2001, Nature Biotechnology.

[34]  Jürgen Weese,et al.  Voxel-based 2-D/3-D registration of fluoroscopy images and CT scans for image-guided surgery , 1997, IEEE Transactions on Information Technology in Biomedicine.

[35]  Shuang Huang,et al.  Elevated urokinase‐specific surface receptor expression is maintained through its interaction with urokinase plasminogen activator , 2007, Molecular carcinogenesis.

[36]  S. Obenauer,et al.  Flat-panel detector-based volume computed tomography: a novel 3D imaging technique to monitor osteolytic bone lesions in a mouse tumor metastasis model. , 2007, Neoplasia.

[37]  Max A. Viergever,et al.  Image registration by maximization of combined mutual information and gradient information , 2000, IEEE Transactions on Medical Imaging.

[38]  B. Gjertsen,et al.  In Vivo Optical Imaging of Acute Myeloid Leukemia by Green Fluorescent Protein: Time-Domain Autofluorescence Decoupling, Fluorophore Quantification, and Localization , 2007, Molecular imaging.