title: Wide-field optical sectioning for live-tissue imaging by plane-projection multiphoton microscopy.
abstract: Optical sectioning provides three-dimensional (3D) information in biological tissues. However, most imaging techniques implemented with optical sectioning are either slow or deleterious to live tissues. Here, we present a simple design for wide-field multiphoton microscopy, which provides optical sectioning at a reasonable frame rate and with a biocompatible laser dosage. The underlying mechanism of optical sectioning is diffuser-based temporal focusing. Axial resolution comparable to confocal microscopy is theoretically derived and experimentally demonstrated. To achieve a reasonable frame rate without increasing the laser power, a low-repetition-rate ultrafast laser amplifier was used in our setup. A frame rate comparable to that of epifluorescence microscopy was demonstrated in the 3D imaging of fluorescent protein expressed in live epithelial cell clusters. In this report, our design displays the potential to be widely used for video-rate live-tissue and embryo imaging with axial resolution comparable to laser scanning microscopy.
authors: California Institute of Technology, Bioengineering, Pasadena, California 91125, USA. jyyu@caltech.edu
authors: Jiun-Yann
authors: JY
authors: Yu
authors: Chun-Hung
authors: CH
authors: Kuo
authors: Daniel B
authors: DB
authors: Holland
authors: Yenyu
authors: Y
authors: Chen
authors: Mingxing
authors: M
authors: Ouyang
authors: Geoffrey A
authors: GA
authors: Blake
authors: Ruben
authors: R
authors: Zadoyan
authors: Chin-Lin
authors: CL
authors: Guo
mesh_terms: D000818:Animals; D002460:Cell Line; D004058:Diffusion; D004285:Dogs; D006651:Histocytochemistry / Q000379:methods; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007834:Lasers; D018613:Microscopy, Confocal / Q000379:methods*; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*