Open AccessIn vivo multiphoton imaging of a diverse array of fluorophores to investigate deep neurovascular structure
Author(s) -
David R. Miller,
Ahmed M. Hassan,
Jeremy W. Jarrett,
Flor A. Medina,
Evan P. Perillo,
Kristen Hagan,
S. M. Shams Kazmi,
Taylor A. Clark,
Colin T. Sullender,
Theresa A. Jones,
Boris V. Zemelman,
Andrew K. Dunn
Publication year - 2017
Publication title -
biomedical optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.362
H-Index - 86
ISSN - 2156-7085
DOI - 10.1364/boe.8.003470
Subject(s) - indocyanine green , laser , materials science , optics , preclinical imaging , microscopy , fluorescence lifetime imaging microscopy , fluorescence , neurovascular bundle , biomedical engineering , in vivo , anatomy , biology , medicine , physics , microbiology and biotechnology
We perform high-resolution, non-invasive, in vivo deep-tissue imaging of the mouse neocortex using multiphoton microscopy with a high repetition rate optical parametric amplifier laser source tunable between λ =1,100 and 1,400 nm. By combining the high repetition rate (511 kHz) and high pulse energy (400 nJ) of our amplifier laser system, we demonstrate imaging of vasculature labeled with Texas Red and Indocyanine Green, and neurons expressing tdTomato and yellow fluorescent protein. We measure the blood flow speed of a single capillary at a depth of 1.2 mm, and image vasculature to a depth of 1.53 mm with fine axial steps (5 μ m) and reasonable acquisition times. The high image quality enabled analysis of vascular morphology at depths to 1.45 mm.