A high quantum yield molecule-protein complex fluorophore for near-infrared II imaging

Fluorescence imaging in the second near-infrared window (NIR-II) allows visualization of deep anatomical features with an unprecedented degree of clarity. NIR-II fluorophores draw from a broad spectrum of materials spanning semiconducting nanomaterials to organic molecular dyes, yet unfortunately all water-soluble organic molecules with >1,000 nm emission suffer from low quantum yields that have limited temporal resolution and penetration depth. Here, we report tailoring the supramolecular assemblies of protein complexes with a sulfonated NIR-II organic dye (CH-4T) to produce a brilliant 110-fold increase in fluorescence, resulting in the highest quantum yield molecular fluorophore thus far. The bright molecular complex allowed for the fastest video-rate imaging in the second NIR window with ∼50-fold reduced exposure times at a fast 50 frames-per-second (FPS) capable of resolving mouse cardiac cycles. In addition, we demonstrate that the NIR-II molecular complexes are superior to clinically approved ICG for lymph node imaging deep within the mouse body.

[1]  J. B. Ifft.  Sedimentation equilibrium of proteins in density gradients. , 1976, Biophysical chemistry.

[2]  A. Eychmüller,et al.  Absolute photoluminescence quantum yields of IR26 and IR-emissive Cd(1-x)Hg(x)Te and PbS quantum dots--method- and material-inherent challenges. , 2015, Nanoscale.

[3]  Shuo Diao,et al.  Fluorescence Imaging In Vivo at Wavelengths beyond 1500 nm. , 2015, Angewandte Chemie.

[4]  S Mordon,et al.  Indocyanine green: physicochemical factors affecting its fluorescence in vivo. , 1998, Microvascular research.

[5]  T. Desmettre,et al.  Fluorescence properties and metabolic features of indocyanine green (ICG) as related to angiography. , 2000, Survey of ophthalmology.

[6]  A. Belcher,et al.  Layer-by-layer assembled fluorescent probes in the second near-infrared window for systemic delivery and detection of ovarian cancer , 2016, Proceedings of the National Academy of Sciences.

[7]  Hak Soo Choi,et al.  Synthesis and in vivo fate of zwitterionic near-infrared fluorophores. , 2011, Angewandte Chemie.

[8]  D. Citterio,et al.  Bright, color-tunable fluorescent dyes in the visible-near-infrared region. , 2008, Journal of the American Chemical Society.

[9]  Yunxia Li,et al.  Highly Fluorescent Ribonuclease-A-Encapsulated Lead Sulfide Quantum Dots for Ultrasensitive Fluorescence in Vivo Imaging in the Second Near-Infrared Window , 2016, Chemistry of materials : a publication of the American Chemical Society.

[10]  Shuo Diao,et al.  Ultrafast fluorescence imaging in vivo with conjugated polymer fluorophores in the second near-infrared window , 2014, Nature Communications.

[11]  Chu Tang,et al.  Novel benzo-bis(1,2,5-thiadiazole) fluorophores for in vivo NIR-II imaging of cancer† †Electronic supplementary information (ESI) available. See DOI: 10.1039/c6sc01561a , 2016, Chemical science.

[12]  Manoj Kumar,et al.  A TICT based NIR-fluorescent probe for human serum albumin: a pre-clinical diagnosis in blood serum. , 2016, Chemical communications.

[13]  Neelkanth M. Bardhan,et al.  Carbon Nanotubes as in vivo Bacterial Probes , 2014, Nature Communications.

[14]  Guosong Hong,et al.  Multifunctional in vivo vascular imaging using near-infrared II fluorescence , 2012, Nature Medicine.

[15]  H. Pal,et al.  Efficient fluorescence enhancement and cooperative binding of an organic dye in a supra-biomolecular host-protein assembly. , 2007, Angewandte Chemie.

[16]  Lihong V. Wang,et al.  Photoacoustic imaging and characterization of the microvasculature. , 2010, Journal of biomedical optics.

[17]  K. Drexhage,et al.  Lifetime, photostability, and chemical structure of IR heptamethine cyanine dyes absorbing beyond 1 μm , 1982 .

[18]  Hein Putter,et al.  Randomized, double-blind comparison of indocyanine green with or without albumin premixing for near-infrared fluorescence imaging of sentinel lymph nodes in breast cancer patients , 2011, Breast Cancer Research and Treatment.

[19]  Nicole M. Iverson,et al.  In Vivo Biosensing Via Tissue Localizable Near Infrared Fluorescent Single Walled Carbon Nanotubes , 2013, Nature nanotechnology.

[20]  Leopold Schmetterer,et al.  Use of laser speckle flowgraphy in ocular blood flow research , 2010, Acta ophthalmologica.

[21]  Shuo Diao,et al.  Through-skull fluorescence imaging of the brain in a new near-infrared window , 2014, Nature Photonics.

[22]  Hak Soo Choi,et al.  Targeted zwitterionic near-infrared fluorophores for improved optical imaging , 2013, Nature Biotechnology.

[23]  H. Dai,et al.  Chirality enriched (12,1) and (11,3) single-walled carbon nanotubes for biological imaging. , 2012, Journal of the American Chemical Society.

[24]  Qinglong Qiao,et al.  A twisted-intramolecular-charge-transfer (TICT) based ratiometric fluorescent thermometer with a mega-Stokes shift and a positive temperature coefficient. , 2014, Chemical communications.

[25]  R. C. Benson,et al.  Fluorescence properties of indocyanine green as related to angiography. , 1978, Physics in medicine and biology.

[26]  Gang Qian,et al.  Band Gap Tunable, Donor−Acceptor−Donor Charge-Transfer Heteroquinoid-Based Chromophores: Near Infrared Photoluminescence and Electroluminescence , 2008 .

[27]  Shuo Diao,et al.  A small-molecule dye for NIR-II imaging. , 2016, Nature materials.

[28]  Zhuang Liu,et al.  A route to brightly fluorescent carbon nanotubes for near-infrared imaging in mice. , 2009, Nature nanotechnology.

[29]  Shuo Diao,et al.  Biological imaging using nanoparticles of small organic molecules with fluorescence emission at wavelengths longer than 1000 nm. , 2013, Angewandte Chemie.

[30]  G. Drummen,et al.  Recent advances in twisted intramolecular charge transfer (TICT) fluorescence and related phenomena in materials chemistry , 2016 .

[31]  Andreas Briel,et al.  Stabilization of indocyanine green by encapsulation within micellar systems. , 2009, Molecular pharmaceutics.

[32]  Shuo Diao,et al.  Traumatic Brain Injury Imaging in the Second Near‐Infrared Window with a Molecular Fluorophore , 2016, Advanced materials.

[33]  Robert R. Alfano,et al.  Deep optical imaging of tissue using the second and third near-infrared spectral windows , 2014, Journal of biomedical optics.

[34]  L. Biczók,et al.  Considerable fluorescence enhancement upon supramolecular complex formation between berberine and p-sulfonated calixarenes , 2006 .

[35]  Dejian Zhou,et al.  Direct water-phase synthesis of lead sulfide quantum dots encapsulated by β-lactoglobulin for in vivo second near infrared window imaging with reduced toxicity. , 2016, Chemical communications.

[36]  Nan Li,et al.  Laser Speckle Contrast Imaging: Theory, Instrumentation and Applications , 2013, IEEE Reviews in Biomedical Engineering.

[37]  J. Lakowicz.  Principles of fluorescence spectroscopy , 1983 .

[38]  W. Rettig.  Photophysical and photochemical switches based on twisted intramolecular charge transfer (TICT) states , 1988 .

[39]  J. Luther,et al.  Absolute Photoluminescence Quantum Yields of IR-26 Dye, PbS, and PbSe Quantum Dots , 2010 .

[40]  Shuo Diao,et al.  Near-Infrared II Fluorescence for Imaging Hindlimb Vessel Regeneration With Dynamic Tissue Perfusion Measurement , 2014, Circulation. Cardiovascular imaging.

[41]  Michael S Strano,et al.  M13 phage-functionalized single-walled carbon nanotubes as nanoprobes for second near-infrared window fluorescence imaging of targeted tumors. , 2012, Nano letters.

[42]  Cornelis J H van de Velde,et al.  Optimization of near-infrared fluorescent sentinel lymph node mapping for vulvar cancer. , 2012, American journal of obstetrics and gynecology.

[43]  Martin Hof,et al.  Fluorescence Spectroscopy in Biology , 2005 .

[44]  M. Hof,et al.  Fluorescence spectroscopy in biology : advanced methods and their applications to membranes, proteins, DNA, and cells , 2005 .

[45]  M. Beard,et al.  PbTe colloidal nanocrystals: synthesis, characterization, and multiple exciton generation. , 2006, Journal of the American Chemical Society.

[46]  Sangeeta N. Bhatia,et al.  Deep, noninvasive imaging and surgical guidance of submillimeter tumors using targeted M13-stabilized single-walled carbon nanotubes , 2014, Proceedings of the National Academy of Sciences.