Large-field high-resolution two-photon digital scanned light-sheet microscopy

Zong, Weijian; Zhao, Jingzhe; Chen, Xuanyang; Lin, Yuan; Ren, Huixia; Zhang, Yunfeng; Fan, Mingming; Zhou, Zhuan; Cheng, Heping; Sun, Yujie; Chen, Liangyi

doi:10.1038/cr.2014.124

Cited by 79 publications

(63 citation statements)

References 10 publications

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“…2PE LSFM has been successfully employed to image not only embryos and their organs (Mahou, Vermot, Beaurepaire, & Supatto, ; Olarte, Andilla, Artigas, & Loza‐Alvarez, ; Olarte et al, ; Truong et al, ; Zong et al, ) but also subcellular structures and mechanism inside embryos (like pancreatic islets (Zong et al, ), time lapse dynamics of mitochondria (Zong et al, )) and deep inside highly scattering environments like mouse brain slices (Lavagnino et al, ) and tumor cell clusters (Fahrbach et al, ). As said, 2PE multicolor imaging is tricky and may potentially lead to cumulative photodamage.…”

Section: Recent Technological Developmentsmentioning

confidence: 99%

“…A recent comparative study shows that 2PE Bessel beams outperform Gaussian beams and 1PE Bessel beams in terms of contrast, resolution and penetration depth. They also offer a large uniform illumination areas at the expense of a somewhat poorer SNR (Andilla Truong et al, 2011;Zong et al, 2014Zong et al, , 2015 but also subcellular structures and mechanism inside embryos (like pancreatic islets (Zong et al, 2014), time lapse dynamics of mitochondria (Zong et al, 2015)) and deep inside highly scattering environments like mouse brain slices (Lavagnino et al, 2016b) and tumor cell clusters (Fahrbach et al, 2013a). As said, 2PE multicolor imaging is tricky and may potentially lead to cumulative photodamage.…”

Section: Deep Into Tissues: 2pe Microscopymentioning

confidence: 99%

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Technical implementations of light sheet microscopy

Zagato

Brans

Smedt

et al. 2018

Microscopy Res & Technique

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Fluorescence-based microscopy is among the most successful methods in biological studies. It played a critical role in the visualization of subcellular structures and in the analysis of complex cellular processes, and it is nowadays commonly employed in genetic and drug screenings. Among the fluorescence-based microscopy techniques, light sheet fluorescence microscopy (LSFM) has shown a quite interesting set of benefits. The technique combines the speed of epi-fluorescence acquisition with the optical sectioning capability typical of confocal microscopes. Its unique configuration allows the excitation of only a thin plane of the sample, thus fast, high resolution imaging deep inside tissues is nowadays achievable. The low peak intensity with which the sample is illuminated diminishes phototoxic effects and decreases photobleaching of fluorophores, ensuring data collection for days with minimal adverse consequences on the sample. It is no surprise that LSFM applications have raised in just few years and the technique has been applied to study a wide variety of samples, from whole organism, to tissues, to cell clusters, and single cells. As a consequence, in recent years numerous set-ups have been developed, each one optimized for the type of sample in use and the requirements of the question at hand. Hereby, we aim to review the most advanced LSFM implementations to assist new LSFM users in the choice of the LSFM set-up that suits their needs best. We also focus on new commercial microscopes and "do-it-yourself" strategies; likewise we review recent designs that allow a swift integration of LSFM on existing microscopes.

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Section: Recent Technological Developmentsmentioning

confidence: 99%

Section: Deep Into Tissues: 2pe Microscopymentioning

confidence: 99%

Technical implementations of light sheet microscopy

Zagato

Brans

Smedt

et al. 2018

Microscopy Res & Technique

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“…Here, we provide detailed methodology for imaging of mitochondrial flashes under physiologically relevant conditions, such as in living cells, intact organs (Langendorff perfused heart), and live animals (skeletal muscles) [10, 16, 29, 30]. This in vivo imaging technique has similarities to methods for measuring other signals such as ROS, pH, and calcium (Ca 2+ ), with their respective fluorescent indicators [11–13, 31–33].…”

Section: Introductionmentioning

confidence: 99%

Mitochondrial flashes: From indicator characterization to in vivo imaging

Wang

Zhang

Cheng

2016

Methods

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Mitochondrion is an organelle critically responsible for energy production and intracellular signaling in eukaryotic cells and its dysfunction often accompanies and contributes to human disease. Superoxide is the primary reactive oxygen species (ROS) produced in mitochondria. In vivo detection of superoxide has been a challenge in biomedical research. Here we describe the methods used to characterize a circularly permuted yellow fluorescent protein (cpYFP) as a biosensor for mitochondrial superoxide and pH dynamics. In vitro characterization reveals the high selectivity of cpYFP to superoxide over other ROS species and its dual sensitivity to pH. Confocal and two-photon imaging in conjunction with transgenic expression of the biosensor cpYFP to the mitochondrial matrix detects mitochondrial flash events in living cells, perfused intact hearts, and live animals. The mitochondrial flashes are discrete and stochastic single mitochondrial events triggered by transient mitochondrial permeability transition (tMPT) and composed of a bursting superoxide signal and a transient alkalization signal. The real-time monitoring of single mitochondrial flashes provides a unique tool to study the integrated dynamism of mitochondrial respiration, ROS production, pH regulation and tMPT kinetics under diverse physiological and pathophysiological conditions.

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“…Two-photon fluorescence microscopy has become a prominent tool for high-resolution in vivo imaging of biological tissues (Periasamy et al, 1999;So et al, 2000). Several studies have demonstrated improved techniques for large field-of-view, three-dimensional (3D) imaging of in vivo spec-Correspondence to: Li-Yueh Hsu, DSc, National Heart, Lung and Blood Institute, National Institutes of Health Bldg 10, Rm B1D416, MSC 1061, 10 Center Drive, Bethesda, MD 20892-1061, U.S.A. Tel: (301) 435-5839; fax: (301) 402-2389; e-mail: lyhsu@mail.nih.gov imens at a great depth (Kobat et al, 2009;Schroeder et al, 2011;Crosignani et al, 2012;Zong et al, 2014;Glancy et al, 2014). However, achieving high spatial fidelity in the obtained biological structures is still a challenge due to the distortion of the point-spread function (PSF) of the microscope, the inhomogeneity of tissue and immersion media (Boutet de Monvel et al, 2001;de Monvel et al, 2003;Von Tiedemann et al, 2006), and most importantly, spatially varying distortion resulting from different specimen structures across the imaging field (Booth et al, 2002;Dong et al, 2003;Arigovindan et al, 2010;Maalouf et al, 2011;Temerinac-Ott et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

A Model‐based approach for microvasculature structure distortion correction in two‐photon fluorescence microscopy images

Dao

Lucotte

Chang

et al. 2015

Journal of Microscopy

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SUMMARY This paper investigates a post-processing approach to correct spatial distortion in two-photon fluorescence microscopy images for vascular network reconstruction. It is aimed at in vivo imaging of large field-of-view, deep-tissue studies of vascular structures. Based on simple geometric modeling of the object-of-interest, a distortion function is directly estimated from the image volume by deconvolution analysis. Such distortion function is then applied to sub volumes of the image stack to adaptively adjust for spatially varying distortion and reduce the image blurring through blind deconvolution. The proposed technique was first evaluated in phantom imaging of fluorescent microspheres that are comparable in size to the underlying capillary vascular structures. The effectiveness of restoring three-dimensional spherical geometry of the microspheres using the estimated distortion function was compared with empirically measured point-spread function. Next, the proposed approach was applied to in vivo vascular imaging of mouse skeletal muscle to reduce the image distortion of the capillary structures. We show that the proposed method effectively improve the image quality and reduce spatially varying distortion that occurs in large field-of-view deep-tissue vascular dataset. The proposed method will help in qualitative interpretation and quantitative analysis of vascular structures from fluorescence microscopy images.

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Large-field high-resolution two-photon digital scanned light-sheet microscopy

Cited by 79 publications

References 10 publications

Technical implementations of light sheet microscopy

Technical implementations of light sheet microscopy

Mitochondrial flashes: From indicator characterization to in vivo imaging

A Model‐based approach for microvasculature structure distortion correction in two‐photon fluorescence microscopy images

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