• Advanced Photonics
  • Vol. 1, Issue 1, 016002 (2019)
Peng Fei1、2、*, Jun Nie1, Juhyun Lee3、4, Yichen Ding3、5, Shuoran Li6, Hao Zhang1, Masaya Hagiwara7、8, Tingting Yu2, Tatiana Segura6, Chih-Ming Ho8, Dan Zhu2, and Tzung K. Hsiai3、5、*
Author Affiliations
  • 1Huazhong University of Science and Technology, School of Optical and Electronic Information, Wuhan, China
  • 2Huazhong University of Science and Technology, Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, Wuhan, China
  • 3University of California, Los Angeles, Department of Bioengineering, Los Angeles, California, United States
  • 4University of Texas at Arlington, Joint Department of Bioengineering of UT Arlington/UT Southwestern, Arlington, Texas, United States
  • 5University of California, Los Angeles, School of Medicine, Los Angeles, California, United States
  • 6University of California, Los Angeles, Chemical and Biomolecular Engineering Department, Los Angeles, California, United States
  • 7Osaka Prefecture University, Nanoscience and Nanotechnology Research Center, Research Organization for the 21st Century, Osaka, Japan
  • 8University of California, Los Angeles, Mechanical and Aerospace Engineering Department, Los Angeles, California, United States
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    A key challenge when imaging whole biomedical specimens is how to quickly obtain massive cellular information over a large field of view (FOV). We report a subvoxel light-sheet microscopy (SLSM) method enabling high-throughput volumetric imaging of mesoscale specimens at cellular resolution. A nonaxial, continuous scanning strategy is developed to rapidly acquire a stack of large-FOV images with three-dimensional (3-D) nanoscale shifts encoded. Then, by adopting a subvoxel-resolving procedure, the SLSM method models these low-resolution, cross-correlated images in the spatial domain and can iteratively recover a 3-D image with improved resolution throughout the sample. This technique can surpass the optical limit of a conventional light-sheet microscope by more than three times, with high acquisition speeds of gigavoxels per minute. By fast reconstruction of 3-D cultured cells, intact organs, and live embryos, SLSM method presents a convenient way to circumvent the trade-off between mapping large-scale tissue (>100 mm3) and observing single cell (~1-μm resolution). It also eliminates the need of complicated mechanical stitching or modulated illumination, using a simple light-sheet setup and fast graphics processing unit-based computation to achieve high-throughput, high-resolution 3-D microscopy, which could be tailored for a wide range of biomedical applications in pathology, histology, neuroscience, etc.

    1 Introduction

    In optical microscopy, high-resolution (HR) volumetric imaging of thick biological specimens is highly desirable for many biomedical applications, such as development biology, tissue pathology, digital histology, and neuroscience. To obtain information on cellular events from the larger organism, e.g., a live embryo, intact tissue, or an organ, spatiotemporal patterns from the micro- to mesoscale must be in toto determined and analyzed.15 Thus, there is a growing need to develop HR, high-throughput imaging methods that can map entire large-volume specimens at high-spatiotemporal resolution.6,7 Recently, light-sheet microscopy (LSM) has emerged as a technique of choice that can image samples with low phototoxicity and at high speed.823 However, similar to conventional epifluorescence methods, LSM remains subject to the fundamental trade-off between high illumination/detection numerical apertures (NAs) and wide imaging fields of view (FOVs). In addition, an accurate digital sampling by the camera is also compromised by the need for large pixel size with high-fluorescence sensitivity. Therefore, the achievable resolution of current LSM systems is often pixel-limited under large FOVs, yielding inadequate optical throughput for digital imaging of mesoscale organisms at the cellular resolution. Tile imaging-based LSM systems have been developed to artificially increase the space-bandwidth product (SBP),24 hence, realizing HR imaging of large specimens.18,2529 Despite the compromised speed induced by repetitive mechanical stitching, the high illumination/detection NA configuration in tile imaging induces increased phototoxicity for increasing sample size and limits fluorescence extraction from deep tissue. In addition, several techniques, such as Fourier ptychographic microscopy,30,31 synthetic aperture microscopy,3235 contact-imaging microscopy,36,37 wavelength scanning microscopy,38 and lens-free digital holography,3941 have recently provided a computational means of reconstructing a wide-FOV, HR image based on a number of low-resolution (LR) frames having certain correlations in the space, frequency, or spectrum domain.4244 However, the majority of these methods target two-dimensional (2-D) bright-field microscopy and are not compatible with volumetric fluorescence imaging of thick samples.

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    Peng Fei, Jun Nie, Juhyun Lee, Yichen Ding, Shuoran Li, Hao Zhang, Masaya Hagiwara, Tingting Yu, Tatiana Segura, Chih-Ming Ho, Dan Zhu, Tzung K. Hsiai. Subvoxel light-sheet microscopy for high-resolution high-throughput volumetric imaging of large biomedical specimens[J]. Advanced Photonics, 2019, 1(1): 016002
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    Category: Research Articles
    Received: Jul. 18, 2018
    Accepted: Oct. 17, 2018
    Published Online: Feb. 18, 2019
    The Author Email: Fei Peng (feipeng@hust.edu.cn), Hsiai Tzung K. (thsiai@mednet.ucla.edu)