Full-color optically-sectioned imaging by wide-field microscopy via deep-learning

Wide-field microscopy (WFM) is broadly used in experimental studies of biological specimens. However, combining the out-of-focus signals with the in-focus plane reduces the signal-to-noise ratio (SNR) and axial resolution of the image. Therefore, structured illumination microscopy (SIM) with white light illumination has been used to obtain full-color 3D images, which can capture high SNR optically-sectioned images with improved axial resolution and natural specimen colors. Nevertheless, this full-color SIM (FCSIM) has a data acquisition burden for 3D-image reconstruction with a shortened depth-offield, especially for thick samples such as insects and large-scale 3D imaging using stitching techniques. In this paper, we propose a deep-learning-based method for full-color WFM (FCWFM), i.e., FC-WFM-Deep, which can reconstruct high-quality full-color 3D images with an extended optical sectioning capability directly from FC-WFM z-stack data. The image quality achievable with this FC-WFM-Deep method is comparable to the FC-SIM method in terms of 3D information and spatial resolution, while the reconstruction data size is 21 times smaller and the in-focus depth is doubled. This technique significantly reduces the 3D data acquisition requirements without losing detail and improves the 3D imaging speed by extracting the optical sectioning in the depth-of-field. This cost-effective and convenient method offers a promising tool to observe high-precision color 3D spatial distributions of biological samples.