Abstract
Near-infrared (NIR) fluorescence imaging is an important imaging technology in deep-tissue biomedical imaging and related researches, due to the low absorption and scattering of NIR excitation and/or emission in biological tissues. Laser scanning confocal microscopy (LSCM) plays a significant role in the family of fluorescence microscopy. Due to the introduction of pinhole, it can provide images with optical sectioning, high signal-to-noise ratio and better spatial resolution. In this study, in order to combine the advantages of these two techniques, we set up a fluorescence microscopic imaging system, which can be named as NIR-LSCM. The system was based on a commercially available confocal microscope, utilizing a NIR laser for excitation and a NIR sensitive detector for signal collection. In addition, NIR fluorescent nanoparticles (NPs) were prepared, and utilized for fluorescence imaging of the ear and brain of living mice based on the NIR-LSCM system. The structure of blood vessels at certain depth could be visualized clearly, because of the high-resolution and large-depth imaging capability of NIR-LSCM.
Similar content being viewed by others
References
Chu, L., Wang, S., Li, K., et al.: Biocompatible near-infrared fluorescent nanoparticles for macro and microscopic in vivo functional bioimaging. Biomed. Opt. Express 5(11), 4076–4088 (2014)
Diev, V.V., Schlenker, C.W., Hanson, K., Zhong, Q., Zimmerman, J.D., Forrest, S.R., et al.: Porphyrins fused with unactivated polycyclic aromatic hydrocarbons. J. Org. Chem. 77(1), 143–159 (2012)
Grutzendler, J., Yang, G., Pan, F., Parkhurst, C.N., Gan, W.B.: Transcranial two-photon imaging of the living mouse brain. Cold Spring Harbor Protoc. 2011(9), prot065474 (2011)
Hardham, A.R.: Confocal microscopy in plant-pathogen interactions. Methods Mol. Biol. 835(835), 295–309 (2012)
Horton, N.G., Wang, K., Demirhan, K., Clark, C.G., Wise, F.W., Schaffer, C.B., Xu, C.: In vivo three-photon microscopy of subcortical structures within an intact mouse brain. Nat. Photonics 7(3), 205–209 (2013)
Huang, X., EI-Sayed, I.H., Qian, W., EI-Sayed, M.A.: Cancer cell imaging and photothermal therapy in the near-infrared region by using gold nanorods. J. Am. Chem. Soc. 128(6), 2115–2120 (2006)
Inglefield, J.R., Schwartz-Bloom, R.D.: Confocal imaging of intracellular chloride in living brain slices: measurement of gabaa receptor activity. J. Neurosci. Methods 75(2), 127–135 (1997)
Kim, J.S., Kim, Y.H., Kim, J.H., Kang, K.W., Tae, E.L., Youn, H., et al.: Development and in vivo imaging of a PET/MRI nanoprobe with enhanced NIR fluorescence by dye encapsulation. Nanomedicine 7(2), 219–229 (2012)
Kim, T.I., Jeong, K.H., Min, K.S.: Verrucous epidermal nevus (VEN) successfully treated with indocyanine green (ICG) photodynamic therapy (PDT). Jaad Case Rep. 1(5), 312–314 (2015)
Liu, B., Li, C., Chen, G., Liu, B., Deng, X., Wei, Y., et al.: Synthesis and optimization of MoS2@Fe3O4-ICG/PT(IV) nanoflowers for MR/IR/PA bioimaging and combined PTT/PDT/chemotherapy triggered by 808 nm laser. Adv. Sci. 4(8), 1600540 (2017)
Luo, T., Huang, P., Gao, G., Shen, G., Fu, S., Cui, D., et al.: Mesoporous silica-coated gold nanorods with embedded indocyanine green for dual mode X-ray CT and NIR fluorescence imaging. Opt. Express 19(18), 17030–17039 (2011)
Ntziachristos, V.: Going deeper than microscopy: the optical imaging frontier in biology. Nat. Methods 7(8), 603–614 (2010)
O’Connell, M.K., Murthy, S., Phan, S., Xu, C., Buchanan, J.A., Spilker, R., et al.: The three-dimensional micro- and nanostructure of the aortic medial lamellar unit measured using 3D confocal & electron microscopy imaging. Matrix Biol. 27(3), 171–181 (2008)
Owens, E.A., Lee, S., Choi, J., Henary, M., Choi, H.S.: NIR fluorescent small molecules for intraoperative imaging. Wiley Interdiscip. Rev. Nanomed. Nanobiotechnol. 7(6), 828–838 (2015)
Ryeom, H.K., Kim, S.H., Kim, J.Y., Kim, H.J., Lee, J.M., Chang, Y.M., et al.: Quantitative evaluation of liver function with MRI using Gd-EOB-DTPA. Korean J. Radiol. 5(4), 231–239 (2004)
Shi Kam, N.W., O’Connell, M., Wisdom, J.A., Dai, H.: Carbon nanotubes as multifunctional biological transporters and near-infrared agents for selective cancer cell destruction. Proc. Natl. Acad. Sci. USA 102(33), 11600–11605 (2005)
Tao, H., Yang, K., Ma, Z., Wan, J., Zhang, Y., Kang, Z., et al.: In vivo NIR fluorescence imaging, biodistribution, and toxicology of photoluminescent carbon dots produced from carbon nanotubes and graphite. Small 8(2), 281–290 (2012)
Treger, J.S., Priest, M.F., Iezzi, R., et al.: Real-time imaging of electrical signals with an infrared fda-approved dye. Biophys. J. 107(6), L09–L12 (2014)
Uh, H., Petoud, S.: Novel antennae for the sensitization of near infrared luminescent lanthanide cations. C. R. Chim. 13(6–7), 668–680 (2010)
Webb, R.H.: Confocal optical microscopy. Rep. Prog. Phys. 59(3), 427–471 (1996)
Weissleder, R.: A clearer vision for in vivo imaging. Nat. Biotechnol. 19(4), 316–317 (2001)
Welsher, K., Liu, Z., Daranciang, D., Dai, H.: Selective probing and imaging of cells with single walled carbon nanotubes as near-infrared fluorescent molecules. Nano Lett. 8(2), 586–590 (2008)
Welsher, K., Liu, Z., Sherlock, S.P., et al.: A route to brightly fluorescent carbon nanotubes for near-infrared imaging in mice. Nat. Nanotechnol. 4(11), 773–780 (2009)
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(22), 8943–8948 (2011)
Yang, F., Murugan, R., Wang, S., Ramakrishna, S.: Electrospinning of nano/micro scale poly (L-lactic acid) aligned fibers and their potential in neural tissue engineering. Biomaterials 26(15), 2603–2610 (2005)
Yodh, A., Chance, B.: Spectroscopy and imaging with diffusing light. Phys. Today 48(3), 34–40 (1995)
Acknowledgements
This work was supported by the Zhejiang Provincial Natural Science Foundation of China (LR17F050001) and the National Natural Science Foundation of China (61735016).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Sun, C., Wang, Y., Zhang, H. et al. Near-infrared laser scanning confocal microscopy and its application in bioimaging. Opt Quant Electron 50, 35 (2018). https://doi.org/10.1007/s11082-017-1309-8
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s11082-017-1309-8