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 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 Automatic Detection Technology of Sonar Image Target Based on the Three-Dimensional Imaging

2017 ◽  
Vol 2017 ◽  
pp. 1-8 ◽  
Author(s):  
Wanzeng Kong ◽  
Jinshuai Yu ◽  
Ying Cheng ◽  
Weihua Cong ◽  
Huanhuan Xue
Keyword(s):  
Data Storage ◽  
3D Imaging ◽  
Signal To Noise Ratio ◽  
Three Dimensional ◽  
Automatic Detection ◽  
Classification Model ◽  
Image Detection ◽  
Practical Applications ◽  
Sonar Image ◽  
Detection Technology

With 3D imaging of the multisonar beam and serious interference of image noise, detecting objects based only on manual operation is inefficient and also not conducive to data storage and maintenance. In this paper, a set of sonar image automatic detection technologies based on 3D imaging is developed to satisfy the actual requirements in sonar image detection. Firstly, preprocessing was conducted to alleviate the noise and then the approximate position of object was obtained by calculating the signal-to-noise ratio of each target. Secondly, the separation of water bodies and strata is realized by maximum variance between clusters (OTSU) since there exist obvious differences between these two areas. Thus image segmentation can be easily implemented on both. Finally, the feature extraction is carried out, and the multidimensional Bayesian classification model is established to do classification. Experimental results show that the sonar-image-detection technology can effectively detect the target and meet the requirements of practical applications.

Cubic spline-based depth-dependent localization of mitochondria–endoplasmic reticulum contacts by three-dimensional light-sheet super-resolution microscopy

The Analyst ◽  
2021 ◽  
Author(s):  
Yucheng Sun ◽  
Seungah Lee ◽  
Seong Ho Kang
Keyword(s):  
Endoplasmic Reticulum ◽  
Calcium Homeostasis ◽  
Three Dimensional ◽  
Super Resolution ◽  
Light Sheet ◽  
Cubic Spline ◽  
Contact Distance ◽  
Super Resolution Microscopy

The contact distance between mitochondria (Mito) and endoplasmic reticulum (ER) has received considerable attention owing to their crucial function in maintaining lipid and calcium homeostasis. Herein, cubic spline algorithm-based depth-dependent...

Combination Light-Sheet Illumination with Super-Resolution Three-Dimensional Fluorescence Microimaging

2018 ◽  
Vol 45 (3) ◽  
pp. 0307006 ◽  
Author(s):  
谢新林 Xie Xinlin ◽  
陈蓉 Chen Rong ◽  
赵宇轩 Zhao Yuxuan ◽  
费鹏 Fei Peng
Keyword(s):  
Three Dimensional ◽  
Super Resolution ◽  
Light Sheet

Three-Dimensional Live Imaging of Filamentous Fungi with Light Sheet-Based Fluorescence Microscopy (LSFM)

2017 ◽  
pp. 19-31 ◽  
Author(s):  
Francesco Pampaloni ◽  
Laura Knuppertz ◽  
Andrea Hamann ◽  
Heinz D. Osiewacz ◽  
Ernst H. K. Stelzer
Keyword(s):  
Fluorescence Microscopy ◽  
Filamentous Fungi ◽  
Three Dimensional ◽  
Light Sheet ◽  
Live Imaging

scholarly journals Three-Dimensional Imaging via Time-Correlated Single-Photon Counting

2020 ◽  
Vol 10 (6) ◽  
pp. 1930
Author(s):  
Chengkun Fu ◽  
Huaibin Zheng ◽  
Gao Wang ◽  
Yu Zhou ◽  
Hui Chen ◽  
...  
Keyword(s):  
3D Imaging ◽  
Single Photon ◽  
Signal To Noise Ratio ◽  
Photon Counting ◽  
Three Dimensional ◽  
Similarity Index ◽  
Signal To Noise ◽  
Single Photon Counting ◽  
Imaging Results ◽  
Noise Ratio

Three-dimensional (3D) imaging under the condition of weak light and low signal-to-noise ratio is a challenging task. In this paper, a 3D imaging scheme based on time-correlated single-photon counting technology is proposed and demonstrated. The 3D imaging scheme, which is composed of a pulsed laser, a scanning mirror, single-photon detectors, and a time-correlated single-photon counting module, employs time-correlated single-photon counting technology for 3D LiDAR (Light Detection and Ranging). Aided by the range-gated technology, experiments show that the proposed scheme can image the object when the signal-to-noise ratio is decreased to −13 dB and improve the structural similarity index of imaging results by 10 times. Then we prove the proposed scheme can image the object in three dimensions with a lateral imaging resolution of 512 × 512 and an axial resolution of 4.2 mm in 6.7 s. At last, a high-resolution 3D reconstruction of an object is also achieved by using the photometric stereo algorithm.

Tissue-culture light sheet fluorescence microscopy (TC-LSFM) allows long-term imaging of three-dimensional cell cultures under controlled conditions

2014 ◽  
Vol 6 (10) ◽  
pp. 988-998 ◽  
Author(s):  
Francesco Pampaloni ◽  
Ulrich Berge ◽  
Anastasios Marmaras ◽  
Peter Horvath ◽  
Ruth Kroschewski ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Fluorescence Microscopy ◽  
Fluorescence Imaging ◽  
Cell Cultures ◽  
Three Dimensional ◽  
Light Sheet ◽  
Controlled Conditions ◽  
Light Sheet Fluorescence Microscopy ◽  
Dimensional Cell

This novel system for the long-term fluorescence imaging of live three-dimensional cultures provides minimal photodamage, control of temperature, CO2, pH, and media flow.

scholarly journals Depth-varying Density and Organization of Chondrocytes in Immature and Mature Bovine Articular Cartilage Assessed by 3D Imaging and Analysis

2005 ◽  
Vol 53 (9) ◽  
pp. 1109-1119 ◽  
Author(s):  
Kyle D. Jadin ◽  
Benjamin L. Wong ◽  
Won C. Bae ◽  
Kelvin W. Li ◽  
Amanda K. Williamson ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
3D Imaging ◽  
Articular Surface ◽  
Three Dimensional ◽  
Neighboring Cell ◽  
3D Images ◽  
Bovine Articular Cartilage ◽  
Stages Of Growth ◽  
Cell Organization ◽  
Heterogeneous Tissue

Articular cartilage is a heterogeneous tissue, with cell density and organization varying with depth from the surface. The objectives of the present study were to establish a method for localizing individual cells in three-dimensional (3D) images of cartilage and quantifying depth-associated variation in cellularity and cell organization at different stages of growth. Accuracy of nucleus localization was high, with 99% sensitivity relative to manual localization. Cellularity (million cells per cm3) decreased from 290, 310, and 150 near the articular surface in fetal, calf, and adult samples, respectively, to 120, 110, and 50 at a depth of 1.0 mm. The distance/angle to the nearest neighboring cell was 7.9 μm/31°, 7.1 μm/31°, and 9.1 μm/31° for cells at the articular surface of fetal, calf, and adult samples, respectively, and increased/decreased to 11.6 μm/31°, 12.0 μm/30°, and 19.2 μm/25° at a depth of 0.7 mm. The methodologies described here may be useful for analyzing the 3D cellular organization of cartilage during growth, maturation, aging, degeneration, and regeneration.

scholarly journals Extending resolution of structured illumination microscopy with sparse deconvolution

2021 ◽  
Author(s):  
Weisong Zhao ◽  
Shiqun Zhao ◽  
Liuju Li ◽  
Xiaoshuai Huang ◽  
Shijia Xing ◽  
...  
Keyword(s):  
Signal To Noise Ratio ◽  
Three Dimensional ◽  
Super Resolution ◽  
Live Cell ◽  
Frame Rate ◽  
Live Cells ◽  
Photon Flux ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Spatiotemporal Resolution

Abstract The spatial resolutions of live-cell super-resolution microscopes are limited by the maximum collected photon flux. Taking advantage of a priori knowledge of the sparsity and continuity of biological structures, we develop a deconvolution algorithm that further extends the resolution of super-resolution microscopes under the same photon budgets by nearly twofold. As a result, sparse structured illumination microscopy (Sparse-SIM) achieves ~60 nm resolution at a 564 Hz frame rate, allowing it to resolve intricate structural intermediates, including small vesicular fusion pores, ring-shaped nuclear pores formed by different nucleoporins, and relative movements between the inner and outer membranes of mitochondria in live cells. Likewise, sparse deconvolution can be used to increase the three-dimensional resolution and contrast of spinning-disc confocal-based SIM (SD-SIM), and operates under conditions with the insufficient signal-to-noise-ratio, all of which allows routine four-color, three-dimensional, ~90 nm resolution live-cell super-resolution imaging. Overall, sparse deconvolution may be a general tool to push the spatiotemporal resolution limits of live-cell fluorescence microscopy.

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