Programmable Colored Illumination Microscopy (PCIM): A practical and flexible optical staining approach for microscopic contrast enhancement |
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| Affiliation: | 1. Smart Computational Imaging Laboratory (SCILab), Nanjing University of Science and Technology, Nanjing, Jiangsu Province 210094, China;2. Jiangsu Key Laboratory of Spectral Imaging & Intelligent Sense, Nanjing University of Science and Technology, Nanjing, Jiangsu Province 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, No. 200 Xiaolingwei Street, Nanjing, Jiangsu Province 210094, China;2. Jiangsu Key Laboratory of Spectral Imaging and Intelligent Sense, Nanjing, Jiangsu Province 210094, China;3. Smart Computational Imaging Laboratory (SCILab), Nanjing University of Science and Technology, Nanjing, Jiangsu Province 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. 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. Jiangsu Key Laboratory of Spectral Imaging & Intelligence Sense, Nanjing University of Science and Technology, Nanjing 210094, China;3. College of Mechatronic Engineering and Automation, National University of Defense Technology, Changsha 410073, China;4. School of Mechanical and Aerospace Engineering, Nanyang Technological University, Singapore 639798, Singapore |
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| Abstract: | Programmable colored illumination microscopy (PCIM) has been proposed as a flexible optical staining technique for microscopic contrast enhancement. In this method, we replace the condenser diaphragm of a conventional microscope with a programmable thin film transistor-liquid crystal display (TFT-LCD). By displaying different patterns on the LCD, numerous established imaging modalities can be realized, such as bright field, dark field, phase contrast, oblique illumination, and Rheinberg illuminations, which conventionally rely on intricate alterations in the respective microscope setups. Furthermore, the ease of modulating both the color and the intensity distribution at the aperture of the condenser opens the possibility to combine multiple microscopic techniques, or even realize completely new methods for optical color contrast staining, such as iridescent dark-field and iridescent phase-contrast imaging. The versatility and effectiveness of PCIM is demonstrated by imaging of several transparent colorless specimens, such as unstained lung cancer cells, diatom, textile fibers, and a cryosection of mouse kidney. Finally, the potentialities of PCIM for RGB-splitting imaging with stained samples are also explored by imaging stained red blood cells and a histological section. |
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| Keywords: | Microscopy Contrast enhancement Programmable illumination Optical staining |
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