Journal
SCIENCE BULLETIN
Volume 64, Issue 6, Pages 410-416Publisher
ELSEVIER
DOI: 10.1016/j.scib.2019.02.019
Keywords
Confocal microscopy; NIR-II fluorescence; AIE dots; In vivo cerebrovascular maging; TCSPC; FLIM imaging
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Funding
- Department of Chemical and Biomolecular Engineering in National University
- National Natural Science Foundation of China [61735016]
- Zhejiang Provincial Natural Science Foundation of China [LR17F050001]
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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 (ALE) dots as NIR-II fluorescent probes, 800 mu m-deep 3D in vivo cerebrovascular imaging of a mouse was obtained, and the spatial resolution at 700 mu m depth could reach 8.78 mu m. 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. (C) 2019 Science China Press. Published by Elsevier B.V. and Science China Press. All rights reserved.
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