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Ex and in vivo characterization of the wavelength‐dependent 3‐photon action cross‐sections of red fluorescent proteins covering the 1700‐nm window
Author(s) -
Liu Hongji,
Wang Jiaqi,
Peng Xiao,
Zhuang Ziwei,
Qiu Ping,
Wang Ke
Publication year - 2018
Publication title -
journal of biophotonics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.877
H-Index - 66
eISSN - 1864-0648
pISSN - 1864-063X
DOI - 10.1002/jbio.201700351
Subject(s) - fluorescence , wavelength , two photon excitation microscopy , excitation , optics , excitation wavelength , in vivo , ex vivo , fluorescence lifetime imaging microscopy , materials science , biophysics , chemistry , physics , biology , microbiology and biotechnology , quantum mechanics
Multiphoton action cross‐sections are the prerequisite for excitation light selection. At the 1700‐nm window suitable for deep‐tissue imaging, wavelength‐dependent 3‐photon action cross‐sections ησ 3 for RFPs are unknown, preventing wavelength selection. Here we demonstrate: (1) ex vivo measurement of wavelength‐dependent ησ 3 for purified RFPs; (2) a multiphoton imaging guided measurement system for in vivo measurement; and (3) in vivo measurement of wavelength‐dependent ησ 3 in RFP labeled cells. These fundamental results will provide guidelines for excitation wavelength selection for 3‐photon fluorescence imaging of RFPs at the 1700‐nm window, and augment the existing database of multiphoton action cross‐sections for fluorophores.