研究目的
To achieve super-resolution and cross-modality transformations in fluorescence microscopy without the need for making any assumptions about or modeling of the image-formation process.
研究成果
The deep learning approach allows for the generation of super-resolution images directly from images acquired on conventional, diffraction-limited microscopes without a priori knowledge about the sample and/or the image formation process. This approach democratizes super-resolution microscopy and offers benefits such as rapidly imaging larger FOVs and DOFs, creating higher-resolution images with fewer frames and/or lower light doses.
研究不足
The performance of the neural-network-based super-resolution approach is limited by noise, similar to all other super-resolution imaging modalities. The need for accurate alignment and registration between lower-resolution and higher-resolution images is crucial.
1:Experimental Design and Method Selection:
The study employs a generative adversarial network (GAN) model to transform low-resolution images into high-resolution ones using matched pairs of experimentally acquired images.
2:Sample Selection and Data Sources:
Training data include images from wide-field fluorescence, confocal, and TIRF microscopes.
3:List of Experimental Equipment and Materials:
Includes a standard inverted microscope, motorized stage, various objective lenses, and optical filter sets.
4:Experimental Procedures and Operational Workflow:
Involves multi-stage image registration and alignment process between lower-resolution and corresponding higher-resolution images.
5:Data Analysis Methods:
The success of the super-resolution approach is quantified through spatial frequency spectrum analysis and PSF characterization.
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FluoCells Prepared Slide
#2, #1, #3
Thermo Fisher Scientific
Used for imaging multi-labeled bovine pulmonary artery endothelial cells (BPAECs) and other samples.
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Inverted Microscope
IX83
Olympus Life Science
Used for capturing fluorescence microscopy images.
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Objective Lens
UPLSAPO10X2, UPLSAPO20X
Olympus Life Science
Used for acquiring low-resolution and high-resolution images.
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Optical Filter Sets
OSFI3-TXRED-4040C, OSFI3-FITC-2024B, OSFI3-DAPI-5060C
Semrock
Used for imaging different labeled cell structures and organelles.
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Fluorescence Light Source
U-HGLGPS
Olympus Life Science
Used as a light source for fluorescence microscopy.
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sCMOS Camera
ORCA-flash4.0 v2
Hamamatsu Photonics K.K.
Used for recording images.
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Confocal Microscope
TCS SP8
Leica Microsystems
Used for confocal and STED microscopy.
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Objective Lens
HC PL APO 100x/1.40-NA Oil STED White
Leica Microsystems
Used for confocal and STED imaging.
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Hybrid Photodetector
HyD SMD
Leica Microsystems
Used for capturing emission signal.
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TIRF Microscope
Axio Observer
ZEISS
Used for TIRF microscopy.
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sCMOS Camera
ORCA-Flash4.0
Hamamatsu
Used for capturing TIRF images.
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Objective Lens
100x/1.49-NA
Olympus Life Science
Used for TIRF-SIM imaging.
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Motorized Stage
Olympus Life Science
Used for scanning microscope slides.
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