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Proceedings Paper

Polymer dots enable deep in vivo multiphoton fluorescence imaging of cerebrovascular architecture
Author(s): Ahmed M. Hassan; Xu Wu; Jeremy W. Jarrett; Shihan Xu; David R. Miller; Jiangbo Yu; Evan P. Perillo; Yen-Liang Liu; Daniel T. Chiu; Hsin-Chih Yeh; Andrew K. Dunn
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Paper Abstract

Deep in vivo imaging of vasculature requires small, bright, and photostable fluorophores suitable for multiphoton microscopy (MPM). Although semiconducting polymer dots (pdots) are an emerging class of highly fluorescent contrast agents with favorable advantages for the next generation of in vivo imaging, their use for deep multiphoton imaging has never before been demonstrated. Here we characterize the multiphoton properties of three pdot variants (CNPPV, PFBT, and PFPV) and demonstrate deep imaging of cortical microvasculature in C57 mice. Specifically, we measure the two- versus three-photon power dependence of these pdots and observe a clear three-photon excitation signature at wavelengths longer than 1300 nm, and a transition from two-photon to three-photon excitation within a 1060 – 1300 nm excitation range. Furthermore, we show that pdots enable in vivo two-photon imaging of cerebrovascular architecture in mice up to 850 μm beneath the pial surface using 800 nm excitation. In contrast with traditional multiphoton probes, we also demonstrate that the broad multiphoton absorption spectrum of pdots permits imaging at longer wavelengths (λex = 1,060 and 1225 nm). These wavelengths approach an ideal biological imaging wavelength near 1,300 nm and confer compatibility with a high-power ytterbium-fiber laser and a high pulse energy optical parametric amplifier, resulting in substantial improvements in signal-to-background ratio (>3.5-fold) and greater cortical imaging depths of 900 μm and 1300 μm. Ultimately, pdots are a versatile tool for MPM due to their extraordinary brightness and broad absorption, which will undoubtedly unlock the ability to interrogate deep structures in vivo.

Paper Details

Date Published: 23 February 2018
PDF: 15 pages
Proc. SPIE 10498, Multiphoton Microscopy in the Biomedical Sciences XVIII, 104982Y (23 February 2018); doi: 10.1117/12.2290404
Show Author Affiliations
Ahmed M. Hassan, The Univ. of Texas at Austin (United States)
Xu Wu, Univ. of Washington (United States)
Jeremy W. Jarrett, The Univ. of Texas at Austin (United States)
Shihan Xu, Univ. of Washington (United States)
David R. Miller, The Univ. of Texas at Austin (United States)
Jiangbo Yu, Univ. of Washington (United States)
Evan P. Perillo, The Univ. of Texas at Austin (United States)
Yen-Liang Liu, The Univ. of Texas at Austin (United States)
Daniel T. Chiu, Univ. of Washington (United States)
Hsin-Chih Yeh, The Univ. of Texas at Austin (United States)
Andrew K. Dunn, The Univ. of Texas at Austin (United States)

Published in SPIE Proceedings Vol. 10498:
Multiphoton Microscopy in the Biomedical Sciences XVIII
Ammasi Periasamy; Peter T. C. So; Karsten König; Xiaoliang S. Xie, Editor(s)

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