NIR-II ?uorescence in vivo confocal microscopy with aggregation-induced emission dots
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摘要
Significantly reduced tissue scattering of fluorescence signals in the second near-infrared(NIR-Ⅱ,1,000–1,700 nm)spectral region offers opportunities for large-depth in vivo bioimaging.Nowadays,most reported works concerning NIR-II fluorescence in vivo bioimaging are realized by wide-field illumination and 2D-arrayed detection(e.g.,via InGaAs camera),which has high temporal resolution but limited spatial resolution due to out-of-focus signals.Combining NIR-II fluorescence imaging with confocal microscopy is a good approach to achieve high-spatial resolution visualization of biosamples even at deep tissues.In this presented work,a NIR-II fluorescence confocal microscopic system was setup.By using a kind of aggregation-induced emission(AIE)dots as NIR-II fluorescent probes,800 lm-deep 3D in vivo cerebrovascular imaging of a mouse was obtained,and the spatial resolution at 700 lm depth could reach 8.78 lm.Moreover,the time-correlated single photon counting(TCSPC)technique and femtosecond laser excitation were introduced into NIR-II fluorescence confocal microscopy,and in vivo confocal NIR-II fluorescence lifetime microscopic imaging(FLIM)of mouse cerebral vasculature was successfully realized.
Significantly reduced tissue scattering of fluorescence signals in the second near-infrared(NIR-Ⅱ,1,000–1,700 nm)spectral region offers opportunities for large-depth in vivo bioimaging.Nowadays,most reported works concerning NIR-II fluorescence in vivo bioimaging are realized by wide-field illumination and 2D-arrayed detection(e.g.,via InGaAs camera),which has high temporal resolution but limited spatial resolution due to out-of-focus signals.Combining NIR-II fluorescence imaging with confocal microscopy is a good approach to achieve high-spatial resolution visualization of biosamples even at deep tissues.In this presented work,a NIR-II fluorescence confocal microscopic system was setup.By using a kind of aggregation-induced emission(AIE)dots as NIR-II fluorescent probes,800 lm-deep 3D in vivo cerebrovascular imaging of a mouse was obtained,and the spatial resolution at 700 lm depth could reach 8.78 lm.Moreover,the time-correlated single photon counting(TCSPC)technique and femtosecond laser excitation were introduced into NIR-II fluorescence confocal microscopy,and in vivo confocal NIR-II fluorescence lifetime microscopic imaging(FLIM)of mouse cerebral vasculature was successfully realized.
Significantly reduced tissue scattering of fluorescence signals in the second near-infrared(NIR-Ⅱ,1,000–1,700 nm)spectral region offers opportunities for large-depth in vivo bioimaging.Nowadays,most reported works concerning NIR-II fluorescence in vivo bioimaging are realized by wide-field illumination and 2D-arrayed detection(e.g.,via InGaAs camera),which has high temporal resolution but limited spatial resolution due to out-of-focus signals.Combining NIR-II fluorescence imaging with confocal microscopy is a good approach to achieve high-spatial resolution visualization of biosamples even at deep tissues.In this presented work,a NIR-II fluorescence confocal microscopic system was setup.By using a kind of aggregation-induced emission(AIE)dots as NIR-II fluorescent probes,800 lm-deep 3D in vivo cerebrovascular imaging of a mouse was obtained,and the spatial resolution at 700 lm depth could reach 8.78 lm.Moreover,the time-correlated single photon counting(TCSPC)technique and femtosecond laser excitation were introduced into NIR-II fluorescence confocal microscopy,and in vivo confocal NIR-II fluorescence lifetime microscopic imaging(FLIM)of mouse cerebral vasculature was successfully realized.
引文
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[30] Wan H, Yue JY, Zhu SJ, et al. A bright organic NIR-II nano?uorophore for threedimensional imaging into biological tissues. Nat Commun 2018;9:1171.
[31] Becker W, Bergmann A, Konig K, et al. Picosecond?uorescence lifetime microscopy by TCSPC imaging. Multiphoton Microscopy Biomed Sci2001;1:414–9.
[32] Becker W, Bergmann A, Hink MA, et al. Fluorescence lifetime imaging by timecorrelated single-photon counting. Microsc Res Tech 2004;63:58–66.
[33] Duncan RR, Bergmann A, Cousin MA, et al. Multi-dimensional time-correlated single photon counting(TCSPC)?uorescence lifetime imaging microscopy(FLIM)to detect FRET in cells. J Microsc 2004;215:1–12.
[2] Diao S, Blackburn JL, Hong GS, et al. Fluorescence imaging in vivo at wavelengths beyond 1,500 nm. Angew Chem Int Ed 2015;54:14758–62.
[3] Diao S, Hong GS, Antaris AL, et al. Biological imaging without auto?uorescence in the second near-infrared region. Nano Res 2015;8:3027–34.
[4] Wang R, Li XM, Zhou L, et al. Epitaxial seeded growth of rare-earth nanocrystals with ef?cient 800 nm near-Infrared to 1,525 nm shortwavelength infrared downconversion photoluminescence for in vivo bioimaging. Angew Chem Int Ed 2014;53:12086–90.
[5] Hong GS, Diao S, Chang JL, et al. Through-skull?uorescence imaging of the brain in a new near-infrared window. Nat Photon 2014;8:723–30.
[6] Antaris AL, Chen H, Cheng K, et al. A small-molecule dye for NIR-II imaging. Nat Mater 2016;15:235–42.
[7] Alifu N, Zebibula A, Qi J, et al. Single-molecular near-infrared-ii theranostic systems:ultrastable aggregation-induced emission nanoparticles for longterm tracing and ef?cient photothermal therapy. ACS Nano2018;12:11282–93.
[8] Englhard AS, Palaras A, Volgger V, et al. Confocal laser endomicroscopy in head and neck malignancies using FITC-labelled EpCAM-and EGF-R-antibodies in cell lines and tumor biopsies. J Biophoton 2017;10:1365–76.
[9] Goetz M, Deris I, Vieth M, et al. Near-infrared confocal imaging during minilaparoscopy:a novel rigid endomicroscope with increased imaging plane depth. J Hepatol 2010;53:84–90.
[10] Prasad PN. Introduction to nanomedicine and nanobioengineering. New Jersey:Wiley John&Sons; 2012.
[11] Zhou L, Guang Z, Rusen Y, et al. Muscle-driven in vivo nanogenerator. Adv Mater 2010;22:2534–7.
[12] Zheng Q, Shi B, Fan F, et al. In vivo powering of pacemaker by breathing-driven implanted triboelectric nanogenerator. Adv Mater 2014;26:5851–6.
[13] Zhen Q, Zou Y, Zhang Y, et al. Biodegradable triboelectric nanogenerator as a life-time designed implantable power source. Sci Adv 2016;2:e1501478.
[14] Zebibula A, Alifu N, Xia LQ, et al. Ultrastable and biocompatible NIR-II quantum dots for functional bioimaging. Adv Funct Mater 2018;28:1703451.
[15] Hong GS, Lee JC, Robinson JT, et al. Multifunctional in vivo vascular imaging using near-infrared II?uorescence. Nat Med 2012;18:1841–6.
[16] Naczynski DJ, Tan MC, Zevon M, et al. Rare-earth-doped biological composites as in vivo shortwave infrared reporters. Nat Commun 2013;4:2199.
[17] Hong GS, Zou YP, Antaris AL, et al. Ultrafast?uorescence imaging in vivo with conjugated polymer?uorophores in the second near-infrared window. Nat Commun 2014;5:4206.
[18] Birks JB. Photophysics of aromatic molecules. London:Wiley; 1970.
[19] Luo JD, Xie ZL, Lam JWY, et al. Aggregation-induced emission of 1-methyl-1,2,3,4,5-pentaphenylsilole. Chem Commun 2001;18:1740–1.
[20] Zong L, Gong Y, Yu Y, et al. New perylene diimide derivatives:stable red emission, adjustable property from ACQ to AIE, and good device performance with an EQE value of 4.93%. Sci Bull 2018;63:108–16.
[21] Wang C, Li L, Zhan XJ, et al. Blue AIEgens bearing triphenylethylene peripheral:adjustable intramolecular conjugation and good device performance. Sci Bull2016;61:1746–55.
[22] Qian J, Tang BZ. AIE luminogens for bioimaging and theranostics:from organelles to animals. Chem 2017;3:56–91.
[23] Mei J, Leung NL, Kwok RT, et al. Aggregation-induced emission:together we shine, united we soar! Chem Rev 2015;115:11718–940.
[24] Cai X, Bandla A, Mao D, et al. Biocompatible red?uorescent organic nanoparticles with tunable size and aggregation-induced emission for evaluation of blood-brain barrier damage. Adv Mater 2016;28:8760–5.
[25] Qi J, Sun C, Zebibula A, et al. Real-time and high-resolution bioimaging with bright aggregation-induced emission dots in short-wave infrared region. Adv Mater 2018;30:1706856.
[26] Sheng ZH, Guo B, Hu DH, et al. Bright aggregation-induced-emission dots for targeted synergetic NIR-II?uorescence and NIR-I photoacoustic imaging of orthotopic brain tumors. Adv Mater 2018;30:e1800766.
[27] Qian J, Wang D, Cai FH, et al. Observation of multiphoton-induced?uorescence from graphene oxide nanoparticles and applications in in vivo functional bioimaging. Angew Chem Int Ed 2012;51:10570–5.
[28] Wang SY, Liu YH, Zhang DP, et al. Photoactivation of extracellular-signalregulated kinase signaling in target cells by femtosecond laser. Laser Photon Rev 2018;12:1700137.
[29] He H, Li S, Wang S, et al. Manipulation of cellular light from green?uorescent protein by a femtosecond laser. Nat Photon 2012;6:651–6.
[30] Wan H, Yue JY, Zhu SJ, et al. A bright organic NIR-II nano?uorophore for threedimensional imaging into biological tissues. Nat Commun 2018;9:1171.
[31] Becker W, Bergmann A, Konig K, et al. Picosecond?uorescence lifetime microscopy by TCSPC imaging. Multiphoton Microscopy Biomed Sci2001;1:414–9.
[32] Becker W, Bergmann A, Hink MA, et al. Fluorescence lifetime imaging by timecorrelated single-photon counting. Microsc Res Tech 2004;63:58–66.
[33] Duncan RR, Bergmann A, Cousin MA, et al. Multi-dimensional time-correlated single photon counting(TCSPC)?uorescence lifetime imaging microscopy(FLIM)to detect FRET in cells. J Microsc 2004;215:1–12.