High-speed wide-field optical-sectioning fluorescence microscopy based on one-shot structured illumination

Abstract

Wide-field fluorescence microscopy (WFFM) is widely adopted in biomedical studies. However, axial resolution in most WFFM is poor due to the absence of optical-sectioning capability. To achieve wide-field optical-sectioning, several methods have been proposed, most of which need at least two images to reconstruct one optical sectioning image. So, the frame rate of current wide-field optical sectioning microscopy is no more than half of that of conventional WFFM, which may not meet the speed requirement of fast biodynamic studies. We introduce a novel high-speed, wide-field optical sectioning method based on local contrast weighting function and two-dimensional Hilbert-Huang transform, in which only one structured image is required to reconstruct an optical sectioning image. In this way, the loss of temporal resolution in conventional wide-field optical sectioning microscopy is compensated. We validated this method with the imaging of mouse brain slices.