Multi-focus microscope with HiLo algorithm for fast 3-D fluorescent imaging

English version

Autoři: Wei Lin ^aff001; Dongping Wang ^aff001; Yunlong Meng ^aff001; Shih-Chi Chen ^aff001
Působiště autorů: Department of Mechanical and Automation Engineering, The Chinese University of Hong Kong, Shatin, Hong Kong ^aff001; Institute of Modern Optics, Tianjin Key Laboratory of Optoelectronic Sensor and Sensing Network Technology, Nankai University, Tianjin, China ^aff002
Vyšlo v časopise: PLoS ONE 14(9)
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pone.0222729

Souhrn

In this paper, we present a new multi-focus microscope (MFM) system based on a phase mask and HiLo algorithm, achieving high-speed (20 volumes per second), high-resolution, low-noise 3-D fluorescent imaging. During imaging, the emissions from the specimen at nine different depths are simultaneously modulated and focused to different regions on a single CCD chip, i.e., the CCD chip is subdivided into nine regions to record images from the different selected depths. Next, HiLo algorithm is applied to remove the background noises and to form clean 3-D images. To visualize larger volumes, the nine layers are scanned axially, realizing fast 3-D imaging. In the imaging experiments, a mouse kidney sample of ~ 60 × 60 × 16 μm³ is visualized with only 10 raw images, demonstrating substantially enhanced resolution and contrast as well as suppressed background noises. The new method will find important applications in 3-D fluorescent imaging, e.g., recording fast dynamic events at multiple depths in vivo.

Klíčová slova:

Biology and life sciences – Physical sciences – Engineering and technology – Research and analysis methods – Computer and information sciences – Mathematics – Simulation and modeling – Anatomy – Medicine and health sciences – Applied mathematics – Algorithms – Physics – Imaging techniques – Equipment – Optical equipment – Cameras – Renal system – Kidneys – Microscopy – Light microscopy – Fluorescence microscopy – Fluorescence imaging – Signal processing – Optical lenses – Optics – Focal planes – Information theory – Background signal noise

Zdroje

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Multi-focus microscope with HiLo algorithm for fast 3-D fluorescent imaging

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