Optical projection tomography (OPT) creates isotropic 3D imaging of tissue. Two approaches exist today: Wide-field OPT illuminates the entire sample and acquires projection images with a camera; Scanning-laser optical tomography (SLOT) generates the projection with a moving laser beam and point detector. SLOT has superior light collecting efficiency than wide-field optical tomography, making it ideal for tissue fluorescence imaging.
Regardless the approach, traditional OPT has to compromise between the resolution and the depth of view. In traditional SLOT, the focused Gaussian beam diverges quickly from the focused plane, making it impossible to achieve high resolution imaging through a large volume specimen. We report using Bessel beam instead of Gaussian beam to perform SLOT. By illuminating samples with a narrow Bessel beam throughout an extended depth, high-resolution projection images can be measured in large volume.
Under Bessel illumination, the projection image contains signal from annular-rings of the Bessel beam. Traditional inverse Radon transform of these projections will result in ringing artifacts in reconstructed imaging. Thus a modified 3D filtered back projection algorithm is developed to perform tomography reconstructing of Bessel-illuminated projection images. The resulting 3D imaging is free of artifact and achieved cellular resolution in extended sample volume.
The system is applied to in-vivo imaging of transgenic Zebrafish embryos. Results prove Bessel SLOT a promising imaging method in development biology research.
Dongli Xu and Leilei Peng, "High resolution multiplexed functional imaging in live embyros
(Conference Presentation)," Proc. SPIE 9700, Design and Quality for Biomedical Technologies IX, 97000U (Presented at SPIE BiOS: February 14, 2016; Published: 26 April 2016); https://doi.org/10.1117/12.2213994.4848651664001.
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