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WIREs Nanomed Nanobiotechnol
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Fluorescent nanotechnology for in vivo imaging

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Abstract Fluorescent imaging in living animals gives an intuitive picture of the dynamic processes in the complex environment within a living being. However, animal tissues present a substantial barrier and are opaque to most wavelengths of visible light. Fluorescent nanoparticles (NPs) with new photophysical characteristics have shown excellent performance for in vivo imaging. Hence, fluorescent NPs have been widely studied and applied for the detection of molecular and biological processes in living animals. In addition, developments in the area of nanotechnology have allowed materials to be used in intact animals for disease detection, diagnosis, drug delivery, and treatment. This review provides information on the different types of fluorescent particles based on nanotechnology, describing their unique individual properties and applications for detecting vital processes in vivo. The development and application of new fluorescent NPs will provide opportunities for in vivo imaging with better penetration, sensitivity, and resolution. This article is categorized under: Diagnostic Tools > in vivo Nanodiagnostics and Imaging
Application of quantum dots (QDs) for in vivo imaging. (a): Images of mouse injected intradermally with 10 pmol of near‐infrared (NIR) QDs in the left paw (Reprinted with permission from Kim et al. (2004). Copyright 2004 Springer Nature). (b): Images of the mouse shown in a 5 min after reinjection with 1% isosulfan blue and exposure of the actual sentinel lymph node. Left, color video; right, NIR fluorescence images (Reprinted with permission from Kim et al. (2004). Copyright 2004 Springer Nature). (c): in vivo QD fluorescence images showing folate‐enhanced tumor targeting of the FL/QD‐TK or L/QD‐TK after tail vein injection into nude mice bearing at different times (Reprinted with permission from Shao et al. (2015). Copyright 2015 Royal Society of Chemistry). (d): Fluorescence intensity at the tumor site for up to 24 h (Reprinted with permission from Shao et al. (2015). Copyright 2015 Royal Society of Chemistry). (e): Graph of relative signal intensity (n¼3, 100% at 1 min) shows that bioluminescence resonance energy transfer (BRET) signal rapidly decreases with time (Reprinted with permission from Kosaka et al. (2011). Copyright 2011 John Wiley and Sons). (f): Relative signal intensity at different time points (Reprinted with permission from Kosaka et al. (2011). Copyright 2011 John Wiley and Sons)
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Near‐infrared (NIR) fluorescent protein for in vivo imaging. (a): CPCiRFP at eight different dilutions were intramuscularly injected into both right and life sides of mouse hindlimb muscles (Reprinted with permission from Wang et al. (2014). Copyright 2014 John Wiley and Sons). (b): Assessment of infrared fluorescence signals showed a good linear correlation (Reprinted with permission from Wang et al. (2014). Copyright 2014 John Wiley and Sons). (c): Comparison of iRFPs with green fluorescent protein (GFP)‐like far‐red FPs as fluorescent probes for deep‐tissue imaging with protein samples located 7.0 mm deep. (Reprinted with permission from Shcherbakova and Verkhusha (2013). Copyright 2013 Springer Nature). (d): Quantification of the signal‐to‐background ratios for the images shown in c (Reprinted with permission from Shcherbakova and Verkhusha (2013). Copyright 2013 Springer Nature). (e,f): Comparisons as in (c,d) but with protein samples located 18.1 mm deep (Reprinted with permission from Shcherbakova and Verkhusha (2013). Copyright 2013 Springer Nature)
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Visible spectrum fluorescent protein for in vivo imaging. (a): External imaging of extensive locoregional and metastatic growth visualized by red fluorescent protein (RFP) in mice (Reprinted with permission from Katz et al. (2003). Copyright 2003 Elsevier). (b): Open imaging of extensive locoregional and metastatic growth is visualized by RFP in mice (Reprinted with permission from Katz et al. (2003). Copyright 2003 Elsevier). (c): The liver was stripped at autopsy and checked for evidence of metastatic disease (Reprinted with permission from Katz et al. (2003). Copyright 2003 Elsevier). (d): Whole‐body image of orthotopically growing HCT 116‐RFP human colon cancer in green fluorescent protein (GFP) nude mouse (Reprinted with permission from Yang et al. (2003). Copyright 2003 National Academy of Sciences, USA). (e): Visualization of the interaction of host dendritic cells and tumor cells in fresh tumor tissue (Reprinted with permission from Yang et al. (2003). Copyright 2003 National Academy of Sciences, USA). (f): Visualization of extensive host lymphocyte infiltration in fresh tumor tissue. (Reprinted with permission from Yang et al. (2003). Copyright 2003 National Academy of Sciences, USA)
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Applications of carbon dots (CDs) for in vivo imaging. (a): Aqueous solution of the PEG1500N‐attached carbon dots excited at 400 nm and photographed through filters of different wavelengths (Reprinted with permission from Sun et al. (2006). Copyright 2006 American Chemical Society). (b): Aqueous solution of PEG1500N‐attached carbon dots excited at the indicated wavelengths and photographed directly (Reprinted with permission from Sun et al. (2006). Copyright 2006 American Chemical Society). (c): Whole‐body distribution of CD‐Asp after injection at different times (Reprinted with permission from Zheng et al. (2015). Copyright 2015 American Chemical Society). (d): Three‐dimensional reconstruction of CD‐Asp distribution in the brain 20 min postinjection (Reprinted with permission from Zheng et al. (2015). Copyright 2015 American Chemical Society). (e): ex vivo imaging of brain 90 min post the injection of CD‐Asp (Reprinted with permission from Zheng et al. (2015). Copyright 2015 American Chemical Society). (f): Synthetic route to CDs (left) and infrared (IR) thermal imaging of mice tumor sites intratumorally injected with CDs at different times following irradiation with a 671 nm laser (right) (Reprinted with permission from Ge et al. (2015). Copyright 2015 John Wiley and Sons). (g): The working mechanism of two‐photon excited near‐infrared (NIR)‐emissive S, Se‐codoped CDs. (Reprinted with permission from Lan et al. (2017). Copyright 2017 from Springer)
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Application of fluorescent/nuclear imaging for in vivo imaging (Reprinted with permission from Chen et al. (2008). Copyright 2008 Springer). (a): The structure of the 1,4,7,10‐tetraazacyclododecane‐N,N′,N,N′‐tetraacetic acid (DOTA)–quantum dot (QD)–vascular endothelial growth factor (VEGF) conjugate. (b): Electrophoresis results for QD and DOTA‐QD‐VEGF in 2% agarose gel. (c): in vivo near‐infrared (NIR) fluorescence imaging of U87MG tumor‐bearing mice at 10, 30, 60, and 90 min p.i. of 200 pmol of DOTA–QD–VEGF and DOTA–QD. (d): Whole‐body coronal positron emission computed tomography (PET) images of U87MG tumor‐bearing mice at 1, 4, 16, and 24 h p.i. of about 300 μCi of 64Cu–DOTA–QD and 64Cu–DOTA–QD–VEGF
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Application of fluorescence/magnetic resonance (MR) for in vivo imaging (Reprinted with permission from Shibu et al. (2013). Copyright 2013 American Chemical Society). (a): Scheme for photouncaging reaction of substituted coumarins under 400 nm excitation and temporal evolution of UV–vis absorption spectra under photoactivation at 400 nm. (b): Fluorescence and overlay images of B16 cells labeled with photouncaging bimodal nanoparticles (PUNP)‐allatostatin conjugates and Syto 21 dye. (c): Fluorescence and overlay images of H1650 cells labeled with quantum dot (QD)–Regulated Growth Factor(EGF) conjugates and Syto 21 dye. (d): T1‐weighted MR of a B6 mouse subcutaneously injected with PUNP. (e,f): Fluorescence and magnetic resonance imaging (MRI) images of the mouse 24 h after intravenous injection of PUNP
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Application of upconversion nanoparticles (NPs) for in vivo imaging. (a): Schematic diagram showing upconversionfluorescent nanoparticle (UCN)‐based targeted photodynamic therapy (PDT) in a mice model of melanoma intravenously injected with UCNs surface modified with folic acid (FA) and polyethylene glycol (PEG) moieties (Reprinted with permission from Idris et al. (2012). Copyright 2012 Springer Nature) (Reprinted with permission from Idris et al., 2012). (b): Representative photographs of a mice from each group 1–3 intravenously injected with FA‐PEG‐UCNs, unmodified UCNs or phosphate buffer saline (PBS) showing the change in tumor development (highlighted by dashed white circles) before (0 day) and 7 days after PDT treatment (Reprinted with permission from Idris et al. (2012), Copyright 2012 with permission from Springer Nature). (c): Schematic illustration of the characterization of upconversion nanoparticles (UCNPs)–Pt(N3)2(NH3)(py)(CCH2CH2COOH)2 (DPP)–PEG nanoparticles (Reprinted with permission from Dai et al. (2013). Copyright 2013 American Chemical Society). (d,e,f): in vivo imaging of a tumor‐bearing Balb/c mice after injection of NPs at the tumor site upconversion luminescence (d), bright field (e), and overlay images (f) (Reprinted with permission from Dai et al. (2013). Copyright 2013 American Chemical Society)
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(a): Emission bands generated under near‐infrared (NIR) excitation by upconversion nanoparticles (UCNPs) doped with different activator ions. The transitions between energy levels that lead each specific emission bands are indicated. (Reprinted with permission from Zhou et al. (2015). Copyright 2015 Springer Nature). (b): Schematic representation of the main in vivo biological applications of UCNPs (Reprinted with permission from Zhou et al. (2015). Copyright 2015 Springer Nature)
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Applications of aggregation‐induced emission nanoparticles (NPs) for in vivo imaging. (a): Fluorescence photographs of solutions and suspensions of (upper panel) fluorescence in water/acetone mixtures with different fractions of acetone (fa) and (lower panel) hexaphenylsilole in Tetrahydrofuran (THF)/water mixtures with different fractions of water (fw) (Reprinted with permission from Mei et al. (2014). Copyright 2014 John Wiley and Sons). (b): Aggregation‐induced emission (AIE) NPs with large 3PA cross‐section were used for 3PL in vivo angiography of mice brain, with a depth of 500 μm (Reprinted with permission from Zhang et al. (2017). Copyright 2017 Royal Society of Chemistry). (c): in vivo noninvasive fluorescence imaging of H22‐tumor‐bearing mice after intravenous injection of fluorogen‐loaded bovine serum albumin (BSA) NPs (Reprinted with permission from Qin et al. (2011). Copyright 2011 John Wiley and Sons). (d): Average PL intensities for the tumor tissues of mice treated with the fluorogen‐loaded BSA NPs and the bare fluorogen NPs at different times (Reprinted with permission from Qin et al. (2011). Copyright 2011 John Wiley and Sons). (e): Representative noninvasive bioluminescence images of PLuc‐expressing PC‐3 tumors transplanted in severe combined immune deficency (SCID) mice (Reprinted with permission from Qin et al. (2011). Copyright 2011 from American Chemical Society)
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