In vivo near-infrared (NIR) fluorescence imaging is an emerging biomedical imaging modality for use in both fundamental scientific research and clinical practice. Owing to advances in reducing photon scattering, light absorption and autofluorescence through innovations in the broad 700–1,700 nm NIR window, NIR fluorescence affords high imaging resolution with increasing tissue penetration depths. In this Review, we cover recent progress made on NIR fluorescence imaging in both the 700–900 nm NIR-I and the 1,000–1,700 nm NIR-II windows by highlighting an increasingly developing palette of biocompatible NIR fluorophores that span the entire NIR window and include inorganic nanoparticles, organic macromolecules and small molecules with tunable emission wavelengths. Together with advances in imaging instrumentation allowing for the efficient detection of long-wavelength NIR photons, recently developed NIR fluorophores have fuelled biomedical imaging from contrast-enhanced imaging of anatomical structures and molecular imaging of specific biomarkers to functional imaging of physiological activities, both for preclinical animal studies and clinical diagnostics and interventions.
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Ntziachristos, V., Ripoll, J., Wang, L. H. V. & Weissleder, R. Looking and listening to light: the evolution of whole-body photonic imaging. Nat. Biotechnol. 23, 313–320 (2005).
Hong, G., Diao, S., Antaris, A. L. & Dai, H. Carbon nanomaterials for biological imaging and nanomedicinal therapy. Chem. Rev. 115, 10816–10906 (2015).
Naumova, A. V., Modo, M., Moore, A., Murry, C. E. & Frank, J. A. Clinical imaging in regenerative medicine. Nat. Biotechnol. 32, 804–818 (2014).
Johnsen, S. Hidden in plain sight: the ecology and physiology of organismal transparency. Biol. Bull. 201, 301–318 (2001).
Frangioni, J. V. In vivo near-infrared fluorescence imaging. Curr. Opin. Chem. Biol. 7, 626–634 (2003).
Ntziachristos, V. Going deeper than microscopy: the optical imaging frontier in biology. Nat. Methods 7, 603–614 (2010).
Hoover, E. E. & Squier, J. A. Advances in multiphoton microscopy technology. Nat. Photon. 7, 93–101 (2013).
Legant, W. R. et al. High-density three-dimensional localization microscopy across large volumes. Nat. Methods 13, 359–365 (2016).
Horstmeyer, R., Ruan, H. W. & Yang, C. H. Guidestar-assisted wavefront-shaping methods for focusing light into biological tissue. Nat. Photon. 9, 563–571 (2015).
Fujimoto, J. G. Optical coherence tomography for ultrahigh resolution in vivo imaging. Nat. Biotechnol. 21, 1361–1367 (2003).
Smith, A. M., Mancini, M. C. & Nie, S. M. Bioimaging: second window for in vivo imaging. Nat. Nanotech. 4, 710–711 (2009).
Matsui, A. et al. Real-time intra-operative near-infrared fluorescence identification of the extrahepatic bile ducts using clinically available contrast agents. Surgery 148, 87–95 (2010).
Tanaka, E. et al. Real-time assessment of cardiac perfusion, coronary angiography, and acute intravascular thrombi using dual-channel near-infrared fluorescence imaging. J. Thorac. Cardiov. Sur. 138, 133–140 (2009).
Verbeek, F. P. R. et al. Intraoperative near infrared fluorescence guided identification of the ureters using low dose methylene blue: a first in human experience. J. Urology 190, 574–579 (2013).
Troyan, S. L. et al. The FLARETM intraoperative near-infrared fluorescence imaging system: a first-in-human clinical trial in breast cancer sentinel lymph node mapping. Ann. Surg. Oncol. 16, 2943–2952 (2009).
Ashitate, Y. et al. Simultaneous assessment of luminal integrity and vascular perfusion of the gastrointestinal tract using dual-channel near-infrared fluorescence. Mol. Imaging 11, 301–308 (2012).
Verbeek, F. P. R. et al. Optimization of near-infrared fluorescence cholangiography for open and laparoscopic surgery. Surg. Endosc. 28, 1076–1082 (2014).
van der Vorst, J. R. et al. Near-infrared fluorescence-guided resection of colorectal liver metastases. Cancer 119, 3411–3418 (2013).
Tummers, Q. R. J. G. et al. Intraoperative guidance in parathyroid surgery using near-infrared fluorescence imaging and low-dose methylene blue. Surgery 158, 1323–1330 (2015).
Tummers, Q. R. J. G. et al. First experience on laparoscopic near-infrared fluorescence imaging of hepatic uveal melanoma metastases using indocyanine green. Surg. Innov. 22, 20–25 (2015).
Verbeek, F. P. R. et al. Sentinel lymph node biopsy in vulvar cancer using combined radioactive and fluorescence guidance. Int. J. Gynecol. Cancer 25, 1086–1093 (2015).
Choi, H. S. et al. Targeted zwitterionic near-infrared fluorophores for improved optical imaging. Nat. Biotechnol. 31, 148–153 (2013).
Hyun, H. et al. 700-nm zwitterionic near-infrared fluorophores for dual-channel image-guided surgery. Mol. Imaging. Biol. 18, 52–61 (2016).
Choi, H. S. et al. Synthesis and in vivo fate of zwitterionic near-infrared fluorophores. Angew. Chem. Int. Ed. 50, 6258–6263 (2011).
Wang, Y. G. et al. A nanoparticle-based strategy for the imaging of a broad range of tumours by nonlinear amplification of microenvironment signals. Nat. Mater. 13, 204–212 (2014).
Grossi, M. et al. Lysosome triggered near-infrared fluorescence imaging of cellular trafficking processes in real time. Nat. Commun. 7, 10855 (2016).
Zheng, X. C. et al. Hypoxia-specific ultrasensitive detection of tumours and cancer cells in vivo. Nat. Commun. 6, 5834 (2015).
Becker, A. et al. Receptor-targeted optical imaging of tumours with near-infrared fluorescent ligands. Nat. Biotechnol. 19, 327–331 (2001).
Hyun, H. et al. Phosphonated near-infrared fluorophores for biomedical imaging of bone. Angew. Chem. Int. Ed. 53, 10668–10672 (2014).
Zaheer, A. et al. In vivo near-infrared fluorescence imaging of osteoblastic activity. Nat. Biotechnol. 19, 1148–1154 (2001).
Hyun, H. et al. Cartilage-specific near-infrared fluorophores for biomedical imaging. Angew. Chem. Int. Ed. 54, 8648–8652 (2015).
Wada, H. et al. Pancreas-targeted NIR fluorophores for dual-channel image-guided abdominal surgery. Theranostics 5, 1–11 (2015).
Hyun, H. et al. Structure-inherent targeting of near-infrared fluorophores for parathyroid and thyroid gland imaging. Nat. Med. 21, 192–197 (2015).
Park, M. H. et al. Prototype nerve-specific near-infrared fluorophores. Theranostics 4, 823–833 (2014).
Bhushan, K. R., Misra, P., Liu, F., Mathur, S., Lenkinski, R. E. & Frangioni, J. V. Detection of breast cancer microcalcifications using a dual-modality SPECT/NIR fluorescent probe. J. Am. Chem. Soc. 130, 17648–17649 (2008).
Aikawa, E. et al. Osteogenesis associates with inflammation in early-stage atherosclerosis evaluated by molecular imaging in vivo. Circulation 116, 2841–2850 (2007).
Vinegoni, C. et al. Indocyanine green enables near-infrared fluorescence imaging of lipid-rich, inflamed atherosclerotic plaques. Sci. Transl. Med. 384ra, 45 (2011).
Hintersteiner, M. et al. In vivo detection of amyloid-β deposits by near-infrared imaging using an oxazine-derivative probe. Nat. Biotechnol. 23, 577–583 (2005).
Staderini, M., Martin, M. A., Bolognesi, M. L. & Menendez, J. C. Imaging of β-amyloid plaques by near infrared fluorescent tracers: a new frontier for chemical neuroscience. Chem. Soc. Rev. 44, 1807–1819 (2015).
Watanabe, H. et al. Molecular imaging of β-amyloid plaques with near-infrared boron dipyrromethane (BODIPY)-based fluorescent probes. Mol. Imaging 12, 338–347 (2013).
Kim, E. et al. Optimized near-IR fluorescent agents for in vivo imaging of Btk expression. Bioconjugate Chem. 26, 1513–1518 (2015).
van Veggel, F. C. J. M. Near-infrared quantum dots and their delicate synthesis, challenging characterization, and exciting potential applications. Chem. Mater. 26, 111–122 (2014).
Wu, C. F. & Chiu, D. T. Highly fluorescent semiconducting polymer dots for biology and medicine. Angew. Chem. Int. Ed. 52, 3086–3109 (2013).
So, M. K., Xu, C. J., Loening, A. M., Gambhir, S. S. & Rao, J. H. Self-illuminating quantum dot conjugates for in vivo imaging. Nat. Biotechnol. 24, 339–343 (2006).
So, M. K., Loening, A. M., Gambhir, S. S. & Rao, J. H. Creating self-illuminating quantum dot conjugates. Nat. Protoc. 1, 1160–1164 (2006).
Ma, N., Marshall, A. F. & Rao, J. H. Near-infrared light emitting luciferase via biomineralization. J. Am. Chem. Soc. 132, 6884–6885 (2010).
Kamkaew, A. et al. Quantum dot-NanoLuc bioluminescence resonance energy transfer enables tumour imaging and lymph node mapping in vivo. Chem. Commun. 52, 6997–7000 (2016).
Hasegawa, M., Tsukasaki, Y., Ohyanagi, T. & Jin, T. Bioluminescence resonance energy transfer coupled near-infrared quantum dots using GST-tagged luciferase for in vivo imaging. Chem. Commun. 49, 228–230 (2013).
Yao, H. Q., Zhang, Y., Xiao, F., Xia, Z. Y. & Rao, J. H. Quantum dot/bioluminescence resonance energy transfer based highly sensitive detection of proteases. Angew. Chem. Int. Ed. 46, 4346–4349 (2007).
Xiong, L. Q., Shuhendler, A. J. & Rao, J. H. Self-luminescing BRET-FRET near-infrared dots for in vivo lymph-node mapping and tumour imaging. N at. Commun . 3, 1193 (2012).
Zhang, N., Francis, K. P., Prakash, A. & Ansaldi, D. Enhanced detection of myeloperoxidase activity in deep tissues through luminescent excitation of near-infrared nanoparticles. Nat. Med. 19, 500–505 (2013).
Palner, M., Pu, K. Y., Shao, S. & Rao, J. H. Semiconducting polymer nanoparticles with persistent near-infrared luminescence for in vivo optical imaging. Angew. Chem. Int. Ed. 54, 11477–11480 (2015).
Abdukayum, A., Chen, J. T., Zhao, Q. & Yan, X. P. Functional near infrared-emitting Cr3+/Pr3+ Co-doped zinc gallogermanate persistent luminescent nanoparticles with superlong afterglow for in vivo targeted bioimaging. J. Am. Chem. Soc. 135, 14125–14133 (2013).
Maldiney, T. et al. The in vivo activation of persistent nanophosphors for optical imaging of vascularization, tumours and grafted cells. Nat. Mater. 13, 418–426 (2014).
Yu, M. X., Liu, J. B., Ning, X. H. & Zheng, J. High-contrast noninvasive imaging of kidney clearance kinetics enabled by renal clearable nanofluorophores. Angew. Chem. Int. Ed. 54, 15434–15438 (2015).
Choi, H. S. et al. Renal clearance of quantum dots. Nat. Biotechnol. 25, 1165–1170 (2007).
Choi, H. S. et al. Tissue- and organ-selective biodistribution of NIR fluorescent quantum dots. Nano Lett. 9, 2354–2359 (2009).
Yu, M. X. et al. Noninvasive staging of kidney dysfunction enabled by renal-clearable luminescent gold nanoparticles. Angew. Chem. Int. Ed. 55, 2787–2791 (2016).
Yu, M. X. & Zheng, J. Clearance pathways and tumour targeting of imaging nanoparticles. ACS Nano 9, 6655–6674 (2015).
Park, J. H. et al. Biodegradable luminescent porous silicon nanoparticles for in vivo applications. Nat. Mater. 8, 331–336 (2009).
Gu, L. et al. In vivo time-gated fluorescence imaging with biodegradable luminescent porous silicon nanoparticles. Nat. Commun. 4, 2326 (2013).
Sun, X. M. et al. Nano-graphene oxide for cellular imaging and drug delivery. Nano Res. 1, 203–212 (2008).
Li, Y. J. et al. Surface coating-dependent cytotoxicity and degradation of graphene derivatives: towards the design of non-toxic, degradable nano-graphene. Small 10, 1544–1554 (2014).
Shu, X. K. et al. Mammalian expression of infrared fluorescent proteins engineered from a bacterial phytochrome. Science 324, 804–807 (2009).
Filonov, G. S. et al. Bright and stable near-infrared fluorescent protein for in vivo imaging. Nat. Biotechnol. 29, 757–761 (2011).
Shcherbakova, D. M. & Verkhusha, V. V. Near-infrared fluorescent proteins for multicolor in vivo imaging. Nat. Methods 10, 751–754 (2013).
Krumholz, A., Shcherbakova, D. M., Xia, J., Wang, L. H. V. & Verkhusha, V. V. Multicontrast photoacoustic in vivo imaging using near-infrared fluorescent proteins. Sci. Rep. 4, 3939 (2014).
Piatkevich, K. D., Subach, F. V. & Verkhusha, V. V. Far-red light photoactivatable near-infrared fluorescent proteins engineered from a bacterial phytochrome. Nat. Commun. 4, 2153 (2013).
Piatkevich, K. D., Subach, F. V. & Verkhusha, V. V. Engineering of bacterial phytochromes for near-infrared imaging, sensing, and light-control in mammals. Chem. Soc. Rev. 42, 3441–3452 (2013).
Shcherbakova, D. M., Baloban, M. & Verkhusha, V. V. Near-infrared fluorescent proteins engineered from bacterial phytochromes. Curr. Opin. Chem. Biol. 27, 52–63 (2015).
Shcherbakova, D. M. et al. Bright monomeric near-infrared fluorescent proteins as tags and biosensors for multiscale imaging. Nat. Commun. 7, 12405 (2016).
Yu, D. et al. An improved monomeric infrared fluorescent protein for neuronal and tumour brain imaging. Nat. Commun. 5, 3626 (2014).
Yu, D. et al. A naturally monomeric infrared fluorescent protein for protein labeling in vivo. Nat. Methods 12, 763–765 (2015).
Chu, J. et al. Non-invasive intravital imaging of cellular differentiation with a bright red-excitable fluorescent protein. Nat. Methods 11, 572–578 (2014).
Kaberniuk, A. A., Shemetov, A. A. & Verkhusha, V. V. A bacterial phytochrome-based optogenetic system controllable with near-infrared light. Nat. Methods 13, 591–597 (2016).
Zhu, B. H., Robinson, H., Zhang, S. L., Wu, G. & Sevick-Muraca, E. M. Longitudinal far red gene-reporter imaging of cancer metastasis in preclinical models: a tool for accelerating drug discovery. Biomed. Opt. Express 6, 3346–3351 (2015).
Kuchimaru, T. et al. A luciferin analogue generating near-infrared bioluminescence achieves highly sensitive deep-tissue imaging. Nat. Commun. 7, 11856 (2016).
Englman, R. & Jortner, J. The energy gap law for radiationless transitions in large molecules. Mol. Phys. 18, 145–164 (1970).
Song, X., Chen, Q. & Liu, Z. Recent advances in the development of organic photothermal nano-agents. Nano Res. 8, 340–354 (2015).
Xu, C., Shi, P., Li, M., Ren, J. S. & Qu, X. G. A cytotoxic amyloid oligomer self-triggered and NIR-enhanced amyloidosis therapeutic system. Nano Res. 8, 2431–2444 (2015).
Tardivo, J. P. et al. Methylene blue in photodynamic therapy: from basic mechanisms to clinical applications. Photodiagn. Photodyn. Ther. 2, 175–191 (2005).
Zheng, M. B. et al. Robust ICG theranostic nanoparticles for folate targeted cancer imaging and highly effective photothermal therapy. ACS Appl. Mater. Interfaces 6, 6709–6716 (2014).
Yue, C. X. et al. IR-780 dye loaded tumour targeting theranostic nanoparticles for NIR imaging and photothermal therapy. Biomaterials 34, 6853–6861 (2013).
Zheng, M. B. et al. Single-step assembly of DOX/ICG loaded lipid-polymer nanoparticles for highly effective chemo-photothermal combination therapy. ACS Nano 7, 2056–2067 (2013).
Liu, Y. et al. Theranostic near-infrared fluorescent nanoplatform for imaging and systemic siRNA delivery to metastatic anaplastic thyroid cancer. Proc. Natl Acad. Sci. USA 113, 7750–7755 (2016).
James, N. S. et al. Comparative tumour imaging and PDT efficacy of HPPH conjugated in the mono- and di-forms to various polymethine cyanine dyes: part-2. Theranostics 3, 703–718 (2013).
Wu, X. M. et al. In vivo and in situ tracking cancer chemotherapy by highly photostable NIR fluorescent theranostic prodrug. J. Am. Chem. Soc. 136, 3579–3588 (2014).
Gu, K. Z. et al. Real-time tracking and in vivo visualization of β-galactosidase activity in colorectal tumor with a ratiometric near-infrared fluorescent probe. J. Am. Chem. Soc. 138, 5334–5340 (2016).
Vahrmeijer, A. L., Hutteman, M., van der Vorst, J. R., van de Velde, C. J. H. & Frangioni, J. V. Image-guided cancer surgery using near-infrared fluorescence. Nat. Rev. Clin. Oncol. 10, 507–518 (2013).
Mieog, J. S. D. et al. Toward optimization of imaging system and lymphatic tracer for near-infrared fluorescent sentinel lymph node mapping in breast cancer. Ann. Surg. Oncol. 18, 2483–2491 (2011).
Hutteman, M. 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. Bre ast Cancer Res. Treat. 127, 163–170 (2011).
Frangioni, J. V. Nonprofit foundations for open-source biomedical technology development. Nat. Biotechnol. 30, 928–932 (2012).
Burrows, P. E. et al. Lymphatic abnormalities are associated with RASA1 gene mutations in mouse and man. Proc. Natl Acad. Sci. USA 110, 8621–8626 (2013).
Rosenthal, E. L. et al. Safety and tumour specificity of cetuximab-IRDye800 for surgical navigation in head and neck cancer. Clin. Cancer Res. 21, 3658–3666 (2015).
Korb, M. L. et al. Breast cancer imaging using the near-infrared fluorescent agent, CLR1502. Mol. Imaging 14, http://dx.doi.org/10.2310/7290.2014.00040 (2015).
Warram, J. M. et al. Fluorescence-guided resection of experimental malignant glioma using cetuximab-IRDye 800CW. Brit. J. Neurosurg. 29, 850–858 (2015).
Rosenthal, E. L. et al. Successful translation of fluorescence navigation during oncologic surgery: a consensus report. J. Nucl. Med. 57, 144–150 (2016).
Bunschoten, A. et al. Tailoring fluorescent dyes to optimize a hybrid RGD-tracer. Bioconjugate Chem. 27, 1253–1258 (2016).
van Leeuwen, F. W., Valdés-Olmos, R., Buckle, T. & Vidal-Sicart, S. Hybrid surgical guidance based on the integration of radionuclear and optical technologies. Br. J. Radiol. 20150797 (2016).
van den Berg, N. S. et al. First-in-human evaluation of a hybrid modality that allows combined radio- and (near-infrared) fluorescence tracing during surgery. Eur. J. Nucl. Med. Mol. Imaging 42, 1639-1647 (2015).
Qian, X. M. et al. In vivo tumour targeting and spectroscopic detection with surface-enhanced Raman nanoparticle tags. Nat. Biotechnol. 26, 83–90 (2008).
Kircher, M. F. et al. A brain tumour molecular imaging strategy using a new triple-modality MRI-photoacoustic-Raman nanoparticle. Nat. Med. 18, 829–834 (2012).
Gandra, N. et al. Tunable and amplified Raman gold nanoprobes for effective tracking (TARGET): in vivo sensing and imaging. Nanoscale 8, 8486–8494 (2016).
Garai, E., et al. A real-time clinical endoscopic system for intraluminal, multiplexed imaging of surface-enhanced Raman scattering nanoparticles. PLoS ONE 10, e0123185 (2015).
Wang, Y. et al. Quantitative molecular phenotyping with topically applied SERS nanoparticles for intraoperative guidance of breast cancer lumpectomy. Sci. Rep. 6, 21242 (2016).
Rao, A. M. et al. Diameter-selective Raman scattering from vibrational modes in carbon nanotubes. Science 275, 187–191 (1997).
Liu, Z. et al. Multiplexed five-color molecular imaging of cancer cells and tumour tissues with carbon nanotube Raman tags in the near-infrared. Nano Res. 3, 222–233 (2010).
Liu, Z. et al. In vivo biodistribution and highly efficient tumour targeting of carbon nanotubes in mice. Nat. Nanotech. 2, 47–52 (2007).
Liu, Z. et al. Circulation and long-term fate of functionalized, biocompatible single-walled carbon nanotubes in mice probed by Raman spectroscopy. Proc. Natl Acad. Sci. USA 105, 1410–1415 (2008).
Keren, S. et al. Noninvasive molecular imaging of small living subjects using Raman spectroscopy. Proc. Natl Acad. Sci. USA 105, 5844–5849 (2008).
Zavaleta, C. et al. Noninvasive Raman spectroscopy in living mice for evaluation of tumour targeting with carbon nanotubes. Nano Lett. 8, 2800–2805 (2008).
Wang, C. et al. Protamine functionalized single-walled carbon nanotubes for stem cell labeling and in vivo Raman/magnetic resonance/photoacoustic triple-modal imaging. Adv. Funct. Mater. 22, 2363–2375 (2012).
Gao, Y. P. et al. Multifunctional gold nanostar-based nanocomposite: synthesis and application for noninvasive MR-SERS imaging-guided photothermal ablation. Biomaterials 60, 31–41 (2015).
Robinson, J. T. et al. High performance in vivo near-IR (> 1 μm) imaging and photothermal cancer therapy with carbon nanotubes. Nano Res. 3, 779–793 (2010).
Mohs, A. M. et al. Hand-held spectroscopic device for in vivo and intraoperative tumour detection: contrast enhancement, detection sensitivity, and tissue penetration. Anal. Chem. 82, 9058–9065 (2010).
Bohndiek, S. E. et al. A small animal Raman instrument for rapid, wide-area, spectroscopic imaging. Proc. Natl Acad. Sci. USA 110, 12408–12413 (2013).
Zavaleta, C. L. et al. A Raman-based endoscopic strategy for multiplexed molecular imaging. Proc. Natl Acad. Sci. USA 110, 10062–10063 (2013).
Zavaleta, C. L. et al. Multiplexed imaging of surface enhanced Raman scattering nanotags in living mice using noninvasive Raman spectroscopy. Proc. Natl Acad. Sci. USA 106, 13511–13516 (2009).
Welsher, K. et al. A route to brightly fluorescent carbon nanotubes for near-infrared imaging in mice. Nat. Nanotech. 4, 773–780 (2009).
Hong, G. et al. Through-skull fluorescence imaging of the brain in a new near-infrared window. Nat. Photon. 8, 723–730 (2014).
Diao, S. et al. Fluorescence imaging in vivo at wavelengths beyond 1500 nm. Angew. Chem. Int. Ed. 54, 14758–14762 (2015).
Wozniak, B. & Dera, J. Light Absorption in Sea Water 11–81 (Springer, 2007).
Diao, S. et al. Biological imaging without autofluorescence in the second near-infrared region. Nano Res. 8, 3027–3034 (2015).
Villa, I. et al. 1.3 μm emitting SrF2:Nd3+ nanoparticles for high contrast in vivo imaging in the second biological window. Nano Res. 8, 649–665 (2015).
del Rosal, B., Villa, I., Jaque, D. & Sanz-Rodríguez, F. In vivo autofluorescence in the biological windows: the role of pigmentation. J. Biophoton. 9, 1059–1067 (2016).
Rogalski, A. Infrared Detectors (CRC, 2010).
Hansen, M. P. & Malchow, D. S. Overview of SWIR detectors, cameras, and applications. Proc. SPIE Thermosense XXX 6939 69390l (2008).
O’Connell, M. J. et al. Band gap fluorescence from individual single-walled carbon nanotubes. Science 297, 593–596 (2002).
Welsher, K., Sherlock, S. P. & Dai, H. Deep-tissue anatomical imaging of mice using carbon nanotube fluorophores in the second near-infrared window. Proc. Natl Acad. Sci. USA 108, 8943–8948 (2011).
Liu, Z., Tabakman, S., Welsher, K. & Dai, H. Carbon nanotubes in biology and medicine: in vitro and in vivo detection, imaging and drug delivery. Nano Res. 2, 85–120 (2009).
Diao, S. et al. Chirality enriched (12,1) and (11,3) single-walled carbon nanotubes for biological imaging. J. Am. Chem. Soc. 134, 16971–16974 (2012).
Robinson, J. T. et al. In vivo fluorescence imaging in the second near-infrared window with long circulating carbon nanotubes capable of ultrahigh tumour uptake. J. Am. Chem. Soc. 134, 10664–10669 (2012).
Antaris, A. L. et al. Ultra-low doses of chirality sorted (6,5) carbon nanotubes for simultaneous tumour imaging and photothermal therapy. ACS Nano 7, 3644–3652 (2013).
Hong, G. et al. Multifunctional in vivo vascular imaging using near-infrared II fluorescence. Nat. Med. 18, 1841–1846 (2012).
Hong, G. et al. Near-infrared II fluorescence for imaging hindlimb vessel regeneration with dynamic tissue perfusion measurement. Circ. Cardiovasc. Imag. 7, 517–525 (2014).
Yomogida, Y., Tanaka, T., Zhang, M., Yudasaka, M., Wei, X. & Kataura, H. Industrial-scale separation of high-purity single-chirality single-wall carbon nanotubes for biological imaging.. Nat. Commun. 7, 12056 (2016).
Bartholomeusz, G. et al. In vivo therapeutic silencing of hypoxia-inducible factor 1 alpha (HIF-1α) using single-walled carbon nanotubes noncovalently coated with siRNA. Nano Res. 2, 279–291 (2009).
Blanco, E., Shen, H. & Ferrari, M. Principles of nanoparticle design for overcoming biological barriers to drug delivery. Nat. Biotechnol. 33, 941–951 (2015).
Liang, C. et al. Tumour metastasis inhibition by imaging-guided photothermal therapy with single-walled carbon nanotubes. Adv. Mater. 26, 5646–5652 (2014).
Dang, X. N. et al. Virus-templated self-assembled single-walled carbon nanotubes for highly efficient electron collection in photovoltaic devices. Nat. Nanotech. 6, 377–384 (2011).
Yi, H. J. et al. M13 phage-functionalized single-walled carbon nanotubes as nanoprobes for second near-infrared window fluorescence imaging of targeted tumours. Nano Lett. 12, 1176–1183 (2012).
Bardhan, N. M., Ghosh, D. & Belcher, A. M. Carbon nanotubes as in vivo bacterial probes. Nat. Commun. 5, 4918 (2014).
Ghosh, D. et al. Deep, noninvasive imaging and surgical guidance of submillimeter tumours using targeted M13-stabilized single-walled carbon nanotubes. Proc. Natl Acad. Sci. USA 111, 13948–13953 (2014).
Iverson, N. M. et al. In vivo biosensing via tissue-localizable near-infrared-fluorescent single-walled carbon nanotubes. Nat. Nanotech. 8, 873–880 (2013).
Hong, G. et al. In vivo fluorescence imaging with Ag2S quantum dots in the second near-infrared region. Angew. Chem. Int. Ed. 51, 9818–9821 (2012).
Hu, F. et al. Real-time in vivo visualization of tumour therapy by a near-infrared-II Ag2S quantum dot-based theranostic nanoplatform. Nano Res. 8, 1637–1647 (2015).
Dong, B. H. et al. Facile synthesis of highly photoluminescent Ag2Se quantum dots as a new fluorescent probe in the second near-infrared window for in vivo imaging. Chem. Mater. 25, 2503–2509 (2013).
Tsukasaki, Y. et al. Synthesis and optical properties of emission-tunable PbS/CdS core-shell quantum dots for in vivo fluorescence imaging in the second near-infrared window. RSC Adv. 4, 41164–41171 (2014).
Li, C. Y. et al. In vivo real-time visualization of tissue blood flow and angiogenesis using Ag2S quantum dots in the NIR-II window. Biomaterials 35, 393–400 (2014).
Nakane, Y., Tsukasaki, Y., Sakata, T., Yasuda, H. & Jin, T. Aqueous synthesis of glutathione-coated PbS quantum dots with tunable emission for non-invasive fluorescence imaging in the second near-infrared biological window (1000–1400 nm). Chem. Commun. 49, 7584–7586 (2013).
Chen, G. et al. Tracking of transplanted human mesenchymal stem cells in living mice using near-Infrared Ag2S quantum dots. Adv. Funct. Mater. 24, 2481–2488 (2014).
Tsukasaki, Y. et al. A short-wavelength infrared emitting multimodal probe for non-invasive visualization of phagocyte cell migration in living mice. Chem. Commun. 50, 14356–14359 (2014).
Chen, G. C. et al. In vivo real-time visualization of mesenchymal stem cells tropism for cutaneous regeneration using NIR-II fluorescence imaging. Biomaterials 53, 265–273 (2015).
Li, C. Y. et al. Real-time monitoring surface chemistry-dependent in vivo behaviors of protein nanocages via encapsulating an NIR-II Ag2S quantum dot. ACS Nano 9, 12255–12263 (2015).
Huang, S. et al. Development of NIR-II fluorescence image-guided and pH-responsive nanocapsules for cocktail drug delivery. Nano Res. 8, 1932–1943 (2015).
Zhang, Y. et al. Ag2S quantum dot: a bright and biocompatible fluorescent nanoprobe in the second near-infrared window. ACS Nano 6, 3695–3702 (2012).
Gui, R. J., Wan, A. J., Liu, X. F., Yuan, W. & Jin, H. Water-soluble multidentate polymers compactly coating Ag2S quantum dots with minimized hydrodynamic size and bright emission tunable from red to second near-infrared region. Nanoscale 6, 5467–5473 (2014).
Sasaki, A. et al. Recombinant protein (EGFP-Protein G)-coated PbS quantum dots for in vitro and in vivo dual fluorescence (visible and second-NIR) imaging of breast tumour. Nanoscale 7, 5115–5119 (2015).
Kong, Y. F. et al. Highly fluorescent ribonuclease-A-encapsulated lead sulfide quantum dots for ultrasensitive fluorescence in vivo imaging in the second near-infrared window. Chem. Mater. 28, 3041–3050 (2016).
Cui, F. et al. From two-dimensional metal-organic coordination networks to near-infrared luminescent PbS nanoparticle/layered polymer composite materials. Nano Res. 1, 195–202 (2008).
Naczynski, D. J. et al. Rare-earth-doped biological composites as in vivo shortwave infrared reporters. Nat. Commun. 4, 2199 (2013).
Rocha, U. et al. Neodymium-doped LaF3 nanoparticles for fluorescence bioimaging in the second biological window. Small 10, 1141–1154 (2014).
Zhang, X. W. et al. Magnetic and optical properties of NaGdF4:Nd3+, Yb3+, Tm3+ nanocrystals with upconversion/downconversion luminescence from visible to the near-infrared second window. Nano Res. 8, 636–648 (2015).
Sun, L. D., Dong, H., Zhang, P. Z. & Yan, C. H. Upconversion of rare earth nanomaterials. Annu. Rev. Phys. Chem. 66, 619–642 (2015).
Yu, X. F. et al. Dopant-controlled synthesis of water-soluble hexagonal NaYF4 nanorods with efficient upconversion fluorescence for multicolor bioimaging. Nano Res. 3, 51–60 (2010).
del Rosal, B. et al. Neodymium-based stoichiometric ultrasmall nanoparticles for multifunctional deep-tissue photothermal therapy. Adv. Opt. Mater. 4, 782–789 (2016).
Tao, Z. et al. Biological imaging using nanoparticles of small organic molecules with fluorescence emission at wavelengths longer than 1000 nm. Angew. Chem. Int. Ed. 52, 13002–13006 (2013).
Hong, G. et al. Ultrafast fluorescence imaging in vivo with conjugated polymer fluorophores in the second near-infrared window. Nat. Commun. 5, 4206 (2014).
Antaris, A. L. et al. A small-molecule dye for NIR-II imaging. Nat. Mater. 15, 235–242 (2016).
Zhang, Y. et al. Biodistribution, pharmacokinetics and toxicology of Ag2S near-infrared quantum dots in mice. Biomaterials 34, 3639–3646 (2013).
Dou, L. T., Liu, Y. S., Hong, Z. R., Li, G. & Yang, Y. Low-bandgap near-IR conjugated polymers/molecules for organic electronics. Chem. Rev. 115, 12633–12665 (2015).
Dang, X. N. et al. Layer-by-layer assembled fluorescent probes in the second near-infrared window for systemic delivery and detection of ovarian cancer. Proc. Natl Acad. Sci. USA 113, 5179–5184 (2016).
Mei, J., Leung, N. L. C., Kwok, R. T. K., Lam, J. W. Y. & Tang, B. Z. Aggregation-induced emission: together we shine, united we soar!. Chem. Rev. 115, 11718–11940 (2015).
Zhang, X. D. et al. Traumatic brain injury imaging in the second near-infrared window with a molecular fluorophore. Adv. Mater. 28, 6872–6879 (2016).
Sun, Y. et al. Novel benzo-bis (1,2,5-thiadiazole) fluorophores for in vivo NIR-II imaging of cancer. Chem. Sci. 7, 6203–6207 (2016).
Kwon, S. Y., Jiang, S. N., Zheng, J. H., Choy, H. E. & Min, J. J. Rhodobacter sphaeroides, a novel tumour-targeting bacteria that emits natural near-infrared fluorescence. Microbiol. Immunol. 58, 172–179 (2014).
Albrecht-Buehler, G. Autofluorescence of live purple bacteria in the near infrared. Exp. Cell Res. 236, 43–50 (1997).
Deisseroth, K. Optogenetics: 10 years of microbial opsins in neuroscience. Nat. Neurosci. 18, 1213–1225 (2015).
Chung, K. et al. Structural and molecular interrogation of intact biological systems. Nature 497, 332–337 (2013).
Kasthuri, N. et al. Saturated reconstruction of a volume of neocortex. Cell 162, 648–661 (2015).
Chen, F., Tillberg, P. W. & Boyden, E. S. Expansion microscopy. Science 347, 543–548 (2015).
Cang, H., et al. Ex-STORM: expansion single molecule nanoscopy. Preprint at bioRxivhttp://dx.doi.org/10.1101/049403 (2016).
Lin, C. W., Bachilo, S. M., Vu, M., Beckingham, K. M. & Weisman, R. B. Spectral triangulation: a 3D method for locating single-walled carbon nanotubes in vivo. Nanoscale 8, 10348–10357 (2016).
Cruz, L. J. et al. Targeted nanoparticles for the non-invasive detection of traumatic brain injury by optical imaging and fluorine magnetic resonance imaging. Nano Res. 9, 1276–1289 (2016).
Withana, N. P. et al. Labeling of active proteases in fresh-frozen tissues by topical application of quenched activity-based probes. Nat. Protoc. 11, 184–191 (2016).
Phillips, E. et al. Clinical translation of an ultrasmall inorganic optical-PET imaging nanoparticle probe. Sci. Transl. Med. 6,260ra149 (2014).
Page, M. J. et al. Non-invasive imaging and cellular tracking of pulmonary emboli by near-infrared fluorescence and positron-emission tomography. Nat. Commun. 6, 8448 (2015).
Quan, L. et al. Near-infrared emitting fluorescent BODIPY nanovesicles for in vivo molecular imaging and drug delivery. ACS Appl. Mater. Interfaces 6, 16166–16173 (2014).
Liu, H. Y. et al. Quinoxaline-based polymer dots with ultrabright red to near-infrared fluorescence for in vivo biological imaging. J. Am. Chem. Soc. 137, 10420–10429 (2015).
Pu, K. Y. et al. Phosphorylcholine-coated semiconducting polymer nanoparticles as rapid and efficient labeling agents for in vivo cell tracking. Adv. Healthc. Mater. 3, 1292–1298 (2014).
Zhu, H. et al. Multilayered semiconducting polymer nanoparticles with enhanced NIR fluorescence for molecular imaging in cells, zebrafish and mice. Chem. Sci. 7, 5118–5125 (2016).
Li, K. et al. Photostable fluorescent organic dots with aggregation-induced emission (AIE dots) for noninvasive long-term cell tracing. Sci. Rep. 3, 1150 (2013).
Ding, D. et al. Conjugated polymer amplified far-red/near-infrared fluorescence from nanoparticles with aggregation-induced emission characteristics for targeted in vivo imaging. Adv. Healthc. Mater. 2, 500–507 (2013).
Xie, R. G., Chen, K., Chen, X. Y. & Peng, X. G. InAs/InP/ZnSe core/shell/shell quantum dots as near-infrared emitters: bright, narrow-band, non-cadmium containing, and biocompatible. Nano Res. 1, 457–464 (2008).
del Rosal, B. et al. Infrared-emitting QDs for thermal therapy with real-time subcutaneous temperature feedback. Adv. Funct. Mater. 26, 6060–6068 (2016).
Bashkatov, A. N., Genina, E. A. & Tuchin, V. V. Optical properties of skin, subcutaneous, and muscle tissues: a review. J. Innov. Opt. Health Sci. 4, 9–38 (2011).
Horton, N. G. et al. In vivo three-photon microscopy of subcortical structures within an intact mouse brain. Nat. Photon. 7, 205–209 (2013).
Palczewska, G. et al. Noninvasive two-photon microscopy imaging of mouse retina and retinal pigment epithelium through the pupil of the eye. Nat. Med. 20, 785–789 (2014).
Lakowicz, J. R. Principles of Fluorescence Spectroscopy (Springer, 2013).
Viegas, M. S., Martins, T. C., Seco, F. & do Carmo, A. An improved and cost-effective methodology for the reduction of autofluorescence in direct immunofluorescence studies on formalin-fixed paraffin-embedded tissues. Eur. J. Histochem. 51, 59–66 (2007).
This work was supported by grants from the US National Institute of Health to H.D. (5R01CA135109-02 and 1R01HL127113-01A1), a Cal-BRAIN grant to H.D., a Stanford Graduate Fellowship to G.H. and an NSF Graduate Fellowship to A.L.A.
The authors declare no competing financial interests.
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Hong, G., Antaris, A. & Dai, H. Near-infrared fluorophores for biomedical imaging. Nat Biomed Eng 1, 0010 (2017). https://doi.org/10.1038/s41551-016-0010
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