Article
Multidisciplinary Sciences
Sota Takanezawa, Takashi Saitou, Takeshi Imamura
Summary: The use of two-photon excitation in light-sheet microscopy is limited by its narrow excitation range, but a new illumination optics proposed in this study aims to generate longer extent Bessel beams to address this limitation. A microscope constructed with this technique allows for long-term time-lapse observations and high-speed recording with similar to 2-3-μm axial resolution, expanding the applications of two-photon excitation in light-sheet microscopy.
NATURE COMMUNICATIONS
(2021)
Article
Optics
Yanhong Gan, Zitong Ye, Yubing Han, Ye Ma, Chuankang Li, Qiulan Liu, Wenjie Liu, Cuifang Kuang, Xu Liu
Summary: Light sheet fluorescence microscopy (LSFM) is a promising tool for biological research due to its ability to observe living cells dynamically. However, obtaining optimal image quality in LSFM requires precise alignment between the light sheet and detection focal plane. In this study, we propose a fast and accurate autofocusing method based on deep learning to overcome the challenge of unstable focusing in LSFM with a single shot. Our method is compatible with any light sheet imaging setup using a spatial light modulator. It achieves a predicted root-mean-square error of 0.0942 μm within a range of ± 0.7 μm in a light sheet microscope with a 1.1 numerical aperture detection objective. The neural network architecture we propose has the advantages of small memory size, few training data requirements, and good generalization to untrained sample types.
OPTICS AND LASERS IN ENGINEERING
(2023)
Article
Optics
S. Crombez, P. Leclerc, C. Ray, N. Ducros
Summary: We present a computational light-sheet microscope that enables hyperspectral acquisition at high spectral resolution. By focusing the emitted fluorescence light onto the entrance slit of an imaging spectrometer using a cylindrical lens, and illuminating the specimen with a sequence of structured light patterns to capture the spatial dimension orthogonal to the slit, we demonstrate the feasibility of this method and report the initial results in vivo using hydra specimens labeled with two fluorophores.
Article
Engineering, Electrical & Electronic
Panchun Gu, Zengxin Huang, Mi Ping, Wenshuang Li, Meng Xiang, Xizeng Feng, Dengfeng Kuang
Summary: This study presents a novel method to generate multiplane parallel light sheets with high resolution and low phototoxicity using an isosceles triangular array microstructure. Experimental validation and comparison with simulations were performed to evaluate the imaging quality, showing promising results for volumetric imaging in light sheet fluorescence microscopy.
IEEE JOURNAL OF SELECTED TOPICS IN QUANTUM ELECTRONICS
(2021)
Article
Microscopy
Anita Jannasch, Sven A. Szilagyi, Moritz Burmeister, Q. Tyrell Davis, Gero L. Hermsdorf, Suman De, Erik Schaeffer
Summary: Observing highly dynamic processes inside living cells at the single molecule level is crucial for better understanding of biological systems. However, current imaging techniques face limitations in spatial and temporal resolution, photobleaching, and signal-to-background ratio. To overcome these limitations, a reflected light-sheet microscope with active optics has been developed, enabling fast and high contrast two-color acquisition of 3D images. The microscope demonstrated fast volume scanning of a two-color giant unilamellar vesicle (GUV) hemisphere, as well as imaging and tracking of single lipids in the GUV cap. This enhanced microscope allows fast 3D scanning of artificial membrane systems and potentially live cells with single-molecule sensitivity, providing quantitative and molecular insights into cellular processes.
JOURNAL OF MICROSCOPY
(2022)
Article
Multidisciplinary Sciences
Feifei Wang, Zhuoran Ma, Yeteng Zhong, Felix Salazar, Chun Xu, Fuqiang Ren, Liangqiong Qu, Anna M. Wu, Hongjie Dai
Summary: The study developed a novel near-infrared II structured-illumination light-sheet microscopy for large volumetric three-dimensional imaging of deep tissues in live mammals, with improved spatial resolution and background suppression. This technology provides an additional tool for noninvasive volumetric molecular imaging of immune cells in live mammals.
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
(2021)
Article
Nanoscience & Nanotechnology
Yuan Luo, Ming Lun Tseng, Sunil Vyas, Ting-Yu Hsieh, Jui-Ching Wu, Shang-Yang Chen, Hsiao-Fang Peng, Vin-Cent Su, Tzu-Ting Huang, Hsin Yu Kuo, Cheng Hung Chu, Mu Ku Chen, Jia-Wern Chen, Yu-Chun Chen, Kuang-Yuh Huang, Chieh-Hsiung Kuan, Xu Shi, Hiroaki Misawa, Din Ping Tsai
Summary: Light-sheet fluorescent microscopy (LSFM) is the leading technique for in vivo imaging in the fields of disease, medicine, and cell biology research. This study demonstrates the successful integration of a nanophotonic meta-lens as the illumination component for LSFM, addressing the challenges of high image resolution and optical sectioning. With the meta-lens, the LSFM system complexity is significantly reduced, enabling multicolor fluorescent imaging of live biological specimens with cellular resolution.
Correction
Biochemical Research Methods
Chen LI, Adele Moatti, Xuying Zhang, H. Troy Ghashghaei, Alon Greenbaum
Summary: This note from the publisher corrects the spelling of the fifth author's name in [Biomed. Opt. Express 12, 5214 (2021)].
BIOMEDICAL OPTICS EXPRESS
(2022)
Article
Optics
N. R. Subedi, S. Yaraghi, P. S. Jung, G. Kukal, A. G. McDonald, D. N. Christodoulides, A. E. Vasdekis
Summary: Research shows that digitally scanned Airy beams can improve the throughput rates in Raman imaging by an order of magnitude compared to traditional diffraction-limited beams, achieving high contrast and 1 micron axial resolution for three-dimensional imaging of microparticles. This method also achieves orders of magnitude lower irradiation density than coherent Raman imaging schemes, with faster speed and lower phototoxicity.
Article
Biochemical Research Methods
Tingting Zhu, Jun Nie, Tingting Yu, Dan Zhu, Yanyi Huang, Zaozao Chen, Zhongze Gu, Jiang Tang, Dongyu LI, Peng Fei
Summary: Light sheet microscopy combined with a microchip is an emerging tool in biomedical research that improves efficiency. However, noticeable aberrations induced by the complex refractive indices in the chip limit microchip-enhanced light-sheet microscopy. This study introduces a droplet microchip with a polymer index matched to water, enabling large-scale culture of 3D spheroids and time-lapse imaging with high resolution and throughput.
BIOMEDICAL OPTICS EXPRESS
(2023)
Article
Chemistry, Analytical
Clara Gomez-Cruz, Sonia Laguna, Ariadna Bachiller-Pulido, Cristina Quilez, Marina Canadas-Ortega, Ignacio Albert-Smet, Jorge Ripoll, Arrate Munoz-Barrutia
Summary: This manuscript presents a novel architecture of a single-plane illumination microscopy (SPIM) for high-resolution imaging of live processes at a cellular level. The custom-made microscope overcomes limitations of existing techniques, achieving high-speed acquisition capabilities, low phototoxicity, and low mechanical disturbances, and demonstrating excellent imaging performance in microfluidic devices.
Article
Optics
Anne Stockhausen, Juan Eduardo Rodriguez-Gatica, Jens Schweihoff, Martin Karl Schwarz, Ulrich Kubitscheck
Summary: Common light sheet microscopy has limitations in balancing light sheet width and usable field of view, which can be overcome by using low-diverging Airy beams. However, Airy beams have side lobes that degrade image contrast. In this study, we constructed an Airy beam light sheet microscope and developed a deep learning image deconvolution method to remove side lobe effects. The combination of Airy beam light sheet microscopy and deep learning deconvolution allows for rapid and high-quality imaging of large volumes.
Article
Biochemical Research Methods
Kevin M. Dean, Tonmoy Chakraborty, Stephan Daetwyler, Jinlong Lin, Gerard Garrelts, Ons M'Saad, Hannahmariam T. Mekbib, Fabian F. Voigt, Martina Schaettin, Esther T. Stoeckli, Fritjof Helmchen, Joerg Bewersdorf, Reto Fiolka
Summary: The protocol provides detailed instructions for assembling and operating a versatile variant of light-sheet fluorescence microscopy called axially swept light-sheet microscopy (ASLM), which offers an unparalleled combination of field of view, optical resolution, and optical sectioning. It includes information on the working principle, applications, practical tips, part lists, schematics, and software for instrument control of ASLM.
Review
Optics
Xin Chen, Suyi Zhong, Yiwei Hou, Ruijie Cao, Wenyi Wang, Dong Li, Qionghai Dai, Donghyun Kim, Peng Xi
Summary: Structured illumination microscopy (SIM) has become the standard for next-generation wide-field microscopy, offering ultrahigh imaging speed, superresolution, a large field-of-view, and long-term imaging. Advanced reconstruction algorithms are necessary to unlock the full potential of SIM system hardware. This article introduces the theory and implementation modalities of two SIM algorithms, optical sectioning SIM (OS-SIM) and superresolution SIM (SR-SIM), and provides an overview of existing processing algorithms and the development of reconstruction algorithms for different SIM techniques. Comparison of off-the-shelf SIM systems and future perspectives are also discussed.
LIGHT-SCIENCE & APPLICATIONS
(2023)
Article
Biochemical Research Methods
Petra Paie, Gianmaria Calisesi, Alessia Candeo, Andrea Comi, Federico Sala, Francesco Ceccarelli, Ada De Luigi, Pietro Veglianese, Korbinian Muhlberger, Michael Fokine, Gianluca Valentini, Roberto Osellame, Mark Neil, Andrea Bassi, Francesca Bragheri
Summary: Heterogeneity investigation at the single-cell level is crucial in clinical research. The combination of light sheet fluorescence microscopy and structured illumination microscopy in an optofluidic platform enables high throughput super-resolution imaging, allowing comprehensive evaluation of cellular heterogeneity at high resolution.