The use of quantum dots for analysis of chick CAM vasculature.

Quantum dots (QDs) are fluorescent semiconductor nanocrystals that possess a number of superior fluorescent properties compared to more established organic dyes and fluorescent proteins. As a result, QDs are being studied for use in a wide range of biological applications. We have examined QDs for one such application, visualization of blood vessels of the chick chorioallantoic membrane (CAM), a popular model for studying various aspects of blood vessel development including angiogenesis. Intravitally injected QDs were found to be biocompatible and were kept in circulation over the course of 4 days without any observed deleterious effects. QD vascular residence time was tunable through QD surface chemistry modification. We also found that use of QDs with higher emission wavelengths (>655 nm) virtually eliminated all chick-derived autofluorescence and improved depth-of-field imaging. QDs were compared to FITC-dextrans, a fluorescent dye commonly used for imaging CAM vessels. QDs were found to image vessels as well as or better than FITC-dextrans at 2-3 orders of magnitude lower concentration. We also demonstrated that QDs are fixable with low fluorescence loss and thus can be used in conjunction with histological processing for further sample analysis.

[1]  D. Rifkin,et al.  Biological roles of fibroblast growth factor-2. , 1997, Endocrine reviews.

[2]  D. Ribatti,et al.  Basic fibroblast growth factor-induced angiogenesis in the chick embryo chorioallantoic membrane: an electron microscopy study. , 1997, Microvascular research.

[3]  L. Cohn,et al.  Sentinel lymph node mapping of the pleural space. , 2005, Chest.

[4]  James McBride,et al.  Targeting cell surface receptors with ligand-conjugated nanocrystals. , 2002, Journal of the American Chemical Society.

[5]  J. Folkman.  What is the evidence that tumors are angiogenesis dependent? , 1990, Journal of the National Cancer Institute.

[6]  Setsuo Takatani,et al.  Theoretical Analysis of Diffuse Reflectance from a Two-Layer Tissue Model , 1979, IEEE Transactions on Biomedical Engineering.

[7]  J. Folkman.  Tumor angiogenesis: therapeutic implications. , 1971, The New England journal of medicine.

[8]  Catalina Wong,et al.  Fibrin-based biomaterials to deliver human growth factors , 2003, Thrombosis and Haemostasis.

[9]  M. Bruchez,et al.  Lighting up cells with quantum dots. , 2003, BioTechniques.

[10]  S. Gambhir,et al.  Quantum Dots for Live Cells, in Vivo Imaging, and Diagnostics , 2005, Science.

[11]  D. Balding,et al.  HLA Sequence Polymorphism and the Origin of Humans , 2006 .

[12]  D. Defouw,et al.  Differentiation of endothelial barrier function during normal angiogenesis requires homotypic VE-cadherin adhesion. , 2000, Tissue & cell.

[13]  Nestor Torio Padron,et al.  Engineered adipose tissue supplied by functional microvessels. , 2003, Tissue engineering.

[14]  A Lendlein,et al.  The importance of angiogenesis in the interaction between polymeric biomaterials and surrounding tissue. , 2003, Clinical hemorheology and microcirculation.

[15]  Byron Ballou,et al.  Fluorescence imaging of tumors in vivo. , 2005, Current medicinal chemistry.

[16]  Rajan P. Kulkarni,et al.  Quantum dots are powerful multipurpose vital labeling agents in zebrafish embryos , 2005, Developmental dynamics : an official publication of the American Association of Anatomists.

[17]  W. Durán,et al.  Ontogeny of microvascular permeability to macromolecules in the chick chorioallantoic membrane during normal angiogenesis. , 1995, Microvascular Research.

[18]  D. Tank,et al.  A Miniature Head-Mounted Two-Photon Microscope High-Resolution Brain Imaging in Freely Moving Animals , 2001, Neuron.

[19]  Erkki Ruoslahti,et al.  Nanocrystal targeting in vivo , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[20]  T. Hirano,et al.  Nanogel-quantum dot hybrid nanoparticles for live cell imaging. , 2005, Biochemical and biophysical research communications.

[21]  G. Stark,et al.  Chorioallantoic membrane angiogenesis model for tissue engineering: a new twist on a classic model. , 2003, Tissue engineering.

[22]  F. Moussy,et al.  The chick chorioallantoic membrane as a novel in vivo model for the testing of biomaterials. , 2002, Journal of biomedical materials research.

[23]  S. Nie,et al.  In vivo cancer targeting and imaging with semiconductor quantum dots , 2004, Nature Biotechnology.

[24]  K. Tsukada,et al.  Optical bioimaging: from living tissue to a single molecule: imaging and functional analysis of blood flow in organic microcirculation. , 2003, Journal of pharmacological sciences.

[25]  Borivoj Vojnovic,et al.  Intravital imaging of tumour vascular networks using multi-photon fluorescence microscopy. , 2005, Advanced drug delivery reviews.

[26]  Nasim Akhtar,et al.  Angiogenesis assays: a critical overview. , 2003, Clinical chemistry.

[27]  Byron Ballou,et al.  Noninvasive imaging of quantum dots in mice. , 2004, Bioconjugate chemistry.

[28]  K. Spanel‐Borowski,et al.  The chick chorioallantoic membrane as test system for biocompatible materials , 1989, Research in experimental medicine. Zeitschrift fur die gesamte experimentelle Medizin einschliesslich experimenteller Chirurgie.

[29]  D. Defouw,et al.  The microvascular unit of the 6-day chick chorioallantoic membrane: a fluorescent confocal microscopic and ultrastructural morphometric analysis of endothelial permselectivity. , 1993, Microvascular research.

[30]  M. Bruchez,et al.  Immunofluorescent labeling of cancer marker Her2 and other cellular targets with semiconductor quantum dots , 2003, Nature Biotechnology.

[31]  Masato Yasuhara,et al.  Quantum Dots Targeted to the Assigned Organelle in Living Cells , 2004, Microbiology and immunology.

[32]  V. Chernomordik,et al.  Real time in vivo non-invasive optical imaging using near-infrared fluorescent quantum dots1 , 2005 .

[33]  Fluorescein angiography for the detection of metastases of ovarian tumor in the abdominal cavity, a feasibility pilot , 2004, Lasers in surgery and medicine.

[34]  D. Defouw,et al.  Vascular endothelial growth factor fails to acutely modulate endothelial permeability during early angiogenesis in the chick chorioallantoic membrane. , 2000, Microvascular research.

[35]  L. Cohn,et al.  Sentinel Lymph Node Mapping of the Gastrointestinal Tract by Using Invisible Light , 2006, Annals of Surgical Oncology.

[36]  V. Rizzo,et al.  Microvessels of the chick chorioallantoic membrane uniformly restrict albumin extravasation during angiogenesis and endothelial cytodifferentiation. , 1997, Tissue & cell.

[37]  J. Folkman,et al.  Tumor angiogenesis and metastasis--correlation in invasive breast carcinoma. , 1991, The New England journal of medicine.

[38]  Peptide-conjugated quantum dots: imaging the angiotensin type 1 receptor in living cells. , 2005, Methods in molecular biology.

[39]  H. Ni,et al.  Visualization and quantitation of peroxisomes using fluorescent nanocrystals: treatment of rats and monkeys with fibrates and detection in the liver. , 2004, Toxicological sciences : an official journal of the Society of Toxicology.

[40]  Kenji Yamamoto,et al.  Semiconductor quantum dot/albumin complex is a long-life and highly photostable endosome marker. , 2003, Biochemical and biophysical research communications.

[41]  S. Nie,et al.  Quantum dot bioconjugates for ultrasensitive nonisotopic detection. , 1998, Science.

[42]  S A Tschanz,et al.  Intussusceptive angiogenesis: its role in embryonic vascular network formation. , 2000, Circulation research.

[43]  M. Howarth,et al.  Targeting quantum dots to surface proteins in living cells with biotin ligase. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[44]  J. Post,et al.  Quantum dot ligands provide new insights into erbB/HER receptor–mediated signal transduction , 2004, Nature Biotechnology.

[45]  P. Burri,et al.  Intussusceptive arborization contributes to vascular tree formation in the chick chorio-allantoic membrane , 2000, Anatomy and Embryology.

[46]  T. Mihaljevic,et al.  Near-infrared fluorescent type II quantum dots for sentinel lymph node mapping , 2004, Nature Biotechnology.

[47]  D. Defouw,et al.  Restrictive endothelial barrier function during normal angiogenesis in vivo: partial dependence on tyrosine dephosphorylation of beta-catenin. , 2000, Microvascular research.

[48]  F. Moussy,et al.  Ex ova chick chorioallantoic membrane as a novel in vivo model for testing biosensors. , 2003, Journal of biomedical materials research. Part A.