Optical diffraction tomography microscopy with transport of intensity equation using a light-emitting diode array |
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| Institution: | 1. Smart Computational Imaging (SCI) Laboratory, Nanjing University of Science and Technology, Nanjing, Jiangsu 210094, China;2. Jiangsu Key Laboratory of Spectral Imaging & Intelligent Sense, Nanjing University of Science and Technology, Nanjing, Jiangsu 210094, China;1. Nanjing University of Science and Technology, School of Electronic and Optical Engineering, Xiaolingwei 200#, Nanjing 210094, China;2. Nanjing University of Science and Technology, Jiangsu Key Laboratory of Spectral Imaging & Intelligence Sense, Xiaolingwei 200#, Nanjing 210094, China;1. School of Electronic and Optical Engineering, Nanjing University of Science and Technology, Nanjing, 210094, China;2. Nanjing University of Science and Technology, Jiangsu Key Laboratory of Spectral Imaging & Intelligence Sense, Xiaolingwei 200#, Nanjing, 210094, China;1. Brookhaven National Laboratory - NSLS II, 50 Rutherford Dr. Upton, NY 11973-5000, USA;2. School of Aeronautics and Astronautics, Sichuan University, Chengdu 610065, China;3. Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Shanghai 201800, China;4. University of Chinese Academy of Sciences, Beijing 100049, China;5. Jiangsu Key Laboratory of Spectral Imaging & Intelligence Sense, Nanjing University of Science and Technology, Nanjing 210094, China;1. Jiangsu Key Laboratory of Spectral Imaging & Intelligent Sense, Nanjing University of Science and Technology, Nanjing, Jiangsu Province 210094, China;2. Key Laboratory of Photoelectronic Imaging Technology and System, Ministry of Education of China. Beijing Institute of Technology, Beijing 100081, China |
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| Abstract: | Optical diffraction tomography (ODT) is an effective label-free technique for quantitatively refractive index imaging, which enables long-term monitoring of the internal three-dimensional (3D) structures and molecular composition of biological cells with minimal perturbation. However, existing optical tomographic methods generally rely on interferometric configuration for phase measurement and sophisticated mechanical systems for sample rotation or beam scanning. Thereby, the measurement is suspect to phase error coming from the coherent speckle, environmental vibrations, and mechanical error during data acquisition process. To overcome these limitations, we present a new ODT technique based on non-interferometric phase retrieval and programmable illumination emitting from a light-emitting diode (LED) array. The experimental system is built based on a traditional bright field microscope, with the light source replaced by a programmable LED array, which provides angle-variable quasi-monochromatic illumination with an angular coverage of ±37 degrees in both x and y directions (corresponding to an illumination numerical aperture of ～0.6). Transport of intensity equation (TIE) is utilized to recover the phase at different illumination angles, and the refractive index distribution is reconstructed based on the ODT framework under first Rytov approximation. The missing-cone problem in ODT is addressed by using the iterative non-negative constraint algorithm, and the misalignment of the LED array is further numerically corrected to improve the accuracy of refractive index quantification. Experiments on polystyrene beads and thick biological specimens show that the proposed approach allows accurate refractive index reconstruction while greatly reduced the system complexity and environmental sensitivity compared to conventional interferometric ODT approaches. |
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| Keywords: | Phase retrieval Microscopy Image reconstruction techniques Medical and biological imaging |
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