Open AccessOvercoming tissue scattering in wide-field two-photon imaging by extended detection and computational reconstruction
Author(s) -
Yuanlong Zhang,
Tiankuang Zhou,
Xuemei Hu,
Xinyang Li,
Hao Xie,
F. X. Lu,
Lingjie Kong,
Qionghai Dai
Publication year - 2019
Publication title -
optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.394
H-Index - 271
ISSN - 1094-4087
DOI - 10.1364/oe.27.020117
Subject(s) - optics , scattering , point spread function , microscopy , photon , preclinical imaging , penetration depth , physics , materials science , in vivo , microbiology and biotechnology , biology
Compared to point-scanning multiphoton microscopy, line-scanning temporal focusing microscopy (LTFM) is competitive in high imaging speed while maintaining tight axial confinement. However, considering its wide-field detection mode, LTFM suffers from shallow penetration depth as a result of the crosstalk induced by tissue scattering. In contrast to the spatial filtering based on confocal slit detection, here we propose the extended detection LTFM (ED-LTFM), the first wide-field two-photon imaging technique to extract signals from scattered photons and thus effectively extend the imaging depth. By recording a succession of line-shape excited signals in 2D and reconstructing signals under Hessian regularization, we can push the depth limitation of wide-field imaging in scattering tissues. We validate the concept with numerical simulations, and demonstrate the performance of enhanced imaging depth in in vivo imaging of mouse brains.