scholarly journals Network-enabled efficient image restoration for 3D microscopy of turbid biological specimens

2020 ◽  
Author(s):  
Le Xiao ◽  
Chunyu Fang ◽  
Yarong Wang ◽  
Tingting Yu ◽  
Yuxuan Zhao ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Image Restoration ◽  
Signal To Noise Ratio ◽  
Three Dimensional ◽  
Science Research ◽  
Light Sheet ◽  
Cell Nuclei ◽  
Two Photon ◽  
Photon Excitation ◽  
3D Fluorescence Microscopy

AbstractThough three-dimensional (3D) fluorescence microscopy has been an essential tool for modern life science research, the light scattering by biological specimens fundamentally prevents its more widespread applications in live imaging. We hereby report a deep-learning approach, termed ScatNet, that enables reversion of 3D fluorescence microscopy from high-resolution targets to low-quality, light-scattered measurements, thereby allowing restoration for a single blurred and light-scattered 3D image of deep tissue, with achieving improved resolution and signal-to-noise ratio. Our approach can computationally extend the imaging depth for current 3D fluorescence microscopes, without the addition of complicated optics. Combining ScatNet approach with cutting-edge light-sheet fluorescence microscopy, we demonstrate that the image restoration of cell nuclei in the deep layer of live Drosophila melanogaster embryos at single-cell resolution. Applying our approach to two-photon excitation microscopy, we could improve the signal and resolution of neurons in mouse brain beyond the photon ballistic region.

scholarly journals A Review on Dual-Lens Fluorescence Microscopy for Three-Dimensional Imaging

2020 ◽  
Vol 8 ◽  
Author(s):  
Xiaoyan Li ◽  
Yubing Han ◽  
Wenjie Liu ◽  
Cuifang Kuang ◽  
Xu Liu ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
3D Imaging ◽  
Signal To Noise Ratio ◽  
Three Dimensional ◽  
Super Resolution ◽  
Light Sheet ◽  
Animal Cells ◽  
3D Images ◽  
Single Lens ◽  
Model Animal

Three-dimensional (3D) imaging using dual-lens fluorescence microscopies is popular in observing fluorescently labeled biological samples, such as mammalian/model animal cells, tissues, and embryos. Specifically, dual-lens super-resolution fluorescence microscopy methods using two opposing objective lenses allow significantly higher axial resolution and better signal to noise ratio than traditional single-lens counterparts, and thus distinguish more details in 3D images of fine intracellular structures. For 3D imaging of thick tissues and entire embryos, dual-lens light-sheet fluorescence microscopy methods using two objective lenses, either orthogonal or non-orthogonal, to achieve selective plane illumination, can meet the requirements, and thus can be used to observe embryo development and structures of interest in thick tissues. This review summarizes both dual-lens fluorescence microscopy methods, including their principles, configurations, and 3D imaging applications, providing a guideline for biological laboratories with different 3D imaging needs.

scholarly journals Breaking the limit of 3D fluorescence microscopy by dual-stage-processing network

2018 ◽  
Author(s):  
Hao Zhang ◽  
Yuxuan Zhao ◽  
Chunyu Fang ◽  
Guo Li ◽  
Meng Zhang ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Single Cell ◽  
Science Research ◽  
Light Sheet ◽  
Diffraction Limit ◽  
Acquisition Time ◽  
Cell Organelles ◽  
Volume Rate ◽  
Dual Stage ◽  
3D Fluorescence Microscopy

AbstractAlthough three-dimensional (3D) fluorescence microscopy is an essential tool for life science research, the fundamentally-limited optical throughput, as reflected in the compromise between speed and resolution, so far prevents further movement towards faster, clearer, and higher-throughput applications. We herein report a dual-stage mutual-feedback deep-learning approach that allows gradual reversion of microscopy degradation from high-resolution targets to low-resolution images. Using a single blurred-and-pixelated 3D image as input, our trained network infers a 3D output with notably higher resolution and improved contrast. The performance is better than conventional 1-stage network approaches. It pushes the throughput limit of current 3D fluorescence microscopy in three ways: notably reducing the acquisition time for accurate mapping of large organs, breaking the diffraction limit for imaging subcellular events with faster lower-toxicity measurement, and improving temporal resolution for capturing instantaneous biological processes. Combining our network approach with light-sheet fluorescence microscopy, we demonstrate the imaging of vessels and neurons in the mouse brain at single-cell resolution and with a throughput of 6 minutes for a whole brain. We also image cell organelles beyond the diffraction limit at a 2-Hz volume rate, and map neuronal activities of freely-moving C. elegans at single-cell resolution and 30-Hz volume rate.

scholarly journals A versatile compressed sensing scheme for faster and less phototoxic 3D fluorescence microscopy

2017 ◽  
Author(s):  
Maxime Woringer ◽  
Xavier Darzacq ◽  
Christophe Zimmer ◽  
Mustafa Mir
Keyword(s):  
Fluorescence Microscopy ◽  
Compressed Sensing ◽  
Light Exposure ◽  
Three Dimensional ◽  
Light Sheet ◽  
Computational Imaging ◽  
Acquisition Time ◽  
Trade Offs ◽  
Fold Reduction ◽  
3D Fluorescence Microscopy

AbstractThree-dimensional fluorescence microscopy based on Nyquist sampling of focal planes faces harsh trade-offs between acquisition time, light exposure, and signal-to-noise. We propose a 3D compressed sensing approach that uses temporal modulation of the excitation intensity during axial stage sweeping and can be adapted to fluorescence microscopes without hardware modification. We describe implementations on a lattice light sheet microscope and an epifluorescence microscope, and show that images of beads and biological samples can be reconstructed with a 5-10 fold reduction of light exposure and acquisition time. Our scheme opens a new door towards faster and less damaging 3D fluorescence microscopy.OCIS codes: (110.1758) Computational imaging; (170.2520) Fluorescence microscopy; (170.6900) Three-dimensional microscopy.

scholarly journals Light Sheet Fluorescence Microscopy (LSFM) for Two-Photon Excitation Imaging of Thick Samples

2015 ◽  
Vol 108 (2) ◽  
pp. 326a ◽  
Author(s):  
Giuseppe Sancataldo ◽  
Zeno Lavagnino ◽  
Marta d’Amora ◽  
Francesca Cella Zanacchi ◽  
Alberto Diaspro
Keyword(s):  
Fluorescence Microscopy ◽  
Light Sheet ◽  
Two Photon ◽  
Photon Excitation ◽  
Light Sheet Fluorescence Microscopy ◽  
Two Photon Excitation

Two-photon excitation fluorescence microscopy in living systems

1993 ◽  
Vol 51 ◽  
pp. 154-155 ◽  
Author(s):  
David W. Piston
Keyword(s):  
Confocal Microscopy ◽  
Fluorescence Microscopy ◽  
Singlet State ◽  
Excited Electronic State ◽  
Input Power ◽  
Two Photon ◽  
Simultaneous Absorption ◽  
Photon Excitation ◽  
Very High ◽  
Two Photon Excitation

Two-photon excitation fluorescence microscopy provides attractive advantages over confocal microscopy for three-dimensionally resolved fluorescence imaging. Two-photon excitation arises from the simultaneous absorption of two photons in a single quantitized event whose probability is proportional to the square of the instantaneous intensity. For example, two red photons can cause the transition to an excited electronic state normally reached by absorption in the ultraviolet. In our fluorescence experiments, the final excited state is the same singlet state that is populated during a conventional fluorescence experiment. Thus, the fluorophore exhibits the same emission properties (e.g. wavelength shifts, environmental sensitivity) used in typical biological microscopy studies. In practice, two-photon excitation is made possible by the very high local instantaneous intensity provided by a combination of diffraction-limited focusing of a single laser beam in the microscope and the temporal concentration of 100 femtosecond pulses generated by a mode-locked laser. Resultant peak excitation intensities are 106 times greater than the CW intensities used in confocal microscopy, but the pulse duty cycle of 10−5 maintains the average input power on the order of 10 mW, only slightly greater than the power normally used in confocal microscopy.

Two‐photon excitation fluorescence microscopy using a semiconductor laser

Bioimaging ◽  
1995 ◽  
Vol 3 (2) ◽  
pp. 70-75 ◽  
Author(s):  
Pekka E Hänninen ◽  
Martin Schrader ◽  
Erkki Soini ◽  
Stefan W Hell
Keyword(s):  
Fluorescence Microscopy ◽  
Semiconductor Laser ◽  
Two Photon ◽  
Photon Excitation ◽  
Two Photon Excitation
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