Selection of Quantum Dot Wavelengths for Biomedical Assays and Imaging

Fluorescent semiconductor nanocrystals (quantum dots [QDs]) are hypothesized to be excellent contrast agents for biomedical assays and imaging. A unique property of QDs is that their absorbance increases with increasing separation between excitation and emission wavelengths. Much of the enthusiasm for using QDs in vivo stems from this property, since photon yield should be proportional to the integral of the broadband absorption. In this study, we demonstrate that tissue scatter and absorbance can sometimes offset increasing QD absorption at bluer wavelengths, and counteract this potential advantage. By using a previously validated mathematical model, we explored the effects of tissue absorbance, tissue scatter, wavelength dependence of the scatter, water-to- hemoglobin ratio, and tissue thickness on QD performance. We conclude that when embedded in biological fluids and tissues, QD excitation wavelengths will often be quite constrained, and that excitation and emission wavelengths should be selected carefully based on the particular application. Based on our results, we produced near-infrared QDs optimized for imaging surface vasculature with white light excitation and a silicon CCD camera, and used them to image the coronary vasculature in vivo. Taken together, our data should prove useful in designing fluorescent QD contrast agents optimized for specific biomedical applications.

[1]  Erica Klarreich,et al.  Biologists join the dots , 2001, Nature.

[2]  A Paul Alivisatos,et al.  Sorting fluorescent nanocrystals with DNA. , 2002, Journal of the American Chemical Society.

[3]  D. Drabkin,et al.  Spectrophotometric studies; the crystallographic and optical properties of the hemoglobin of man in comparison with those of other species. , 1946, The Journal of biological chemistry.

[4]  Hedi Mattoussi,et al.  Avidin: a natural bridge for quantum dot-antibody conjugates. , 2002, Journal of the American Chemical Society.

[5]  A. Rogach,et al.  Development of IR-emitting colloidal II-VI quantum-dot materials , 2000, IEEE Journal of Selected Topics in Quantum Electronics.

[6]  A. Koller,et al.  Pharmacologic Inhomogeneity Between the Reactivity of Intramural Coronary Arteries and Arterioles , 2001, Journal of cardiovascular pharmacology.

[7]  W. Semmler,et al.  Receptor-targeted optical imaging of tumors with near-infrared fluorescent ligands , 2001, Nature Biotechnology.

[8]  Uri Banin,et al.  Colloidal chemical synthesis and characterization of InAs nanocrystal quantum dots , 1996 .

[9]  H. Mattoussi,et al.  Conjugation of luminescent quantum dots with antibodies using an engineered adaptor protein to provide new reagents for fluoroimmunoassays. , 2002, Analytical chemistry.

[10]  K. Norris,et al.  A new approach for the estimation of body composition: infrared interactance. , 1984, The American journal of clinical nutrition.

[11]  J. Matthew Mauro,et al.  Self-Assembly of CdSe−ZnS Quantum Dot Bioconjugates Using an Engineered Recombinant Protein , 2000 .

[12]  A. Welch,et al.  A review of the optical properties of biological tissues , 1990 .

[13]  S. Pathak,et al.  Hydroxylated quantum dots as luminescent probes for in situ hybridization. , 2001, Journal of the American Chemical Society.

[14]  H. J. van Staveren,et al.  Light scattering in Intralipid-10% in the wavelength range of 400-1100 nm. , 1991, Applied optics.

[15]  J. Mourant,et al.  Predictions and measurements of scattering and absorption over broad wavelength ranges in tissue phantoms. , 1997, Applied optics.

[16]  R. Anderson,et al.  The optics of human skin. , 1981, The Journal of investigative dermatology.

[17]  S L Jacques,et al.  Light transport in tissue: Accurate expressions for one‐dimensional fluence rate and escape function based upon Monte Carlo simulation , 1996, Lasers in surgery and medicine.

[18]  S. Achilefu,et al.  Novel fluorescent contrast agents for optical imaging of in vivo tumors based on a receptor-targeted dye-peptide conjugate platform. , 2001, Journal of biomedical optics.

[19]  Nikolai Gaponik,et al.  THIOL-CAPPING OF CDTE NANOCRYSTALS: AN ALTERNATIVE TO ORGANOMETALLIC SYNTHETIC ROUTES , 2002 .

[20]  J. Matthew Mauro,et al.  Long-term multiple color imaging of live cells using quantum dot bioconjugates , 2003, Nature Biotechnology.

[21]  Moungi G. Bawendi,et al.  On the Absorption Cross Section of CdSe Nanocrystal Quantum Dots , 2002 .

[22]  U. Banin,et al.  Growth and Properties of Semiconductor Core/Shell Nanocrystals with InAs Cores. , 2001 .

[23]  R. Weissleder A clearer vision for in vivo imaging , 2001, Nature Biotechnology.

[24]  Weilie Zhou,et al.  Synthesis and Properties of Lead Selenide Nanocrystal Solids , 2001 .

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

[26]  S. Nie,et al.  Luminescent quantum dots for multiplexed biological detection and imaging. , 2002, Current opinion in biotechnology.

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

[28]  Jun Q. Lu,et al.  Optical properties of porcine skin dermis between 900 nm and 1500 nm , 2001, Physics in medicine and biology.

[29]  K. H. Drexhage,et al.  Fluorescence quantum yield of oxazine and carbazine laser dyes , 1981 .

[30]  L. Kou,et al.  Refractive indices of water and ice in the 0.65- to 2.5 micrometer spectral range , 1993 .

[31]  I Fridolin,et al.  Optical non-invasive technique for vessel imaging: II. A simplified photon diffusion analysis. , 2000, Physics in medicine and biology.

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

[33]  Vincent Noireaux,et al.  In Vivo Imaging of Quantum Dots Encapsulated in Phospholipid Micelles , 2002, Science.

[34]  Alexander Eychmüller,et al.  Wet chemical synthesis and spectroscopic study of CdHgTe nanocrystals with strong near-infrared luminescence , 2000 .

[35]  Christopher B. Murray,et al.  Colloidal synthesis of nanocrystals and nanocrystal superlattices , 2001, IBM J. Res. Dev..

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

[37]  B. Tromberg,et al.  Sources of absorption and scattering contrast for near-infrared optical mammography. , 2001, Academic radiology.

[38]  Alexander Eychmüller,et al.  Colloidally Prepared HgTe Nanocrystals with Strong Room‐Temperature Infrared Luminescence , 1999 .

[39]  B Chance,et al.  Near‐Infrared Images Using Continuous, Phase‐Modulated, and Pulsed Light with Quantitation of Blood and Blood Oxygenation a , 1998, Annals of the New York Academy of Sciences.

[40]  K. Norris,et al.  Spectrophotometry of human hemoglobin in the midinfrared region , 1997 .

[41]  R. Weissleder,et al.  In vivo imaging of tumors with protease-activated near-infrared fluorescent probes , 1999, Nature Biotechnology.

[42]  Christopher B. Murray,et al.  Synthesis and characterization of nearly monodisperse CdE (E = S, Se, Te) semiconductor nanocrystallites , 2005 .

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

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

[45]  R. Anderson,et al.  TRANSMITTANCE OF NONIONIZING RADIATION IN HUMAN TISSUES * , 1981, Photochemistry and photobiology.

[46]  John V Frangioni,et al.  Functional Near-Infrared Fluorescence Imaging for Cardiac Surgery and Targeted Gene Therapy , 2002, Molecular imaging.