scholarly journals Multidirectional digital scanned light-sheet microscopy enables uniform fluorescence excitation and contrast-enhanced imaging

2018 ◽  
Author(s):  
Adam K. Glaser ◽  
Ye Chen ◽  
Chengbo Yin ◽  
Linpeng Wei ◽  
Lindsey A. Barner ◽  
...  
Keyword(s):  
Scattered Light ◽  
Light Sheet ◽  
Line Detection ◽  
Fluorescence Excitation ◽  
Light Sheet Microscopy ◽  
Contrast Enhanced ◽  
Selective Plane Illumination Microscopy ◽  
Light Sheet Fluorescence Microscopy ◽  
Enhanced Imaging ◽  
Imaging Plane

AbstractLight-sheet fluorescence microscopy (LSFM) has emerged as a powerful method for rapid and optically efficient 3D microscopy. Initial LSFM designs utilized a static sheet of light, termed selective plane illumination microscopy (SPIM), which exhibited shadowing artifacts and deteriorated contrast due to light scattering. These issues have been addressed, in part, by multidirectional selective plane illumination microscopy (mSPIM), in which rotation of the light sheet is used to mitigate shadowing artifacts, and digital scanned light-sheet microscopy (DSLM), in which confocal line detection is used to reject scattered light. Here we present a simple passive multidirectional digital scanned light-sheet microscopy (mDSLM) architecture that combines the benefits of mSPIM and DSLM. By utilizing an elliptical Gaussian beam with increased angular diversity in the imaging plane, mDSLM provides shadow-free contrast-enhanced imaging of fluorescently labeled samples.One Sentence SummaryGlaser et al. describe a light-sheet microscopy architecture that enables passive multidirectional illumination with confocal line detection to enable both uniform fluorescence excitation and contrast-enhanced imaging of fluorescently labeled samples.

scholarly journals Multidirectional digital scanned light-sheet microscopy enables uniform fluorescence excitation and contrast-enhanced imaging

2018 ◽  
Vol 8 (1) ◽  
Author(s):  
Adam K. Glaser ◽  
Ye Chen ◽  
Chengbo Yin ◽  
Linpeng Wei ◽  
Lindsey A. Barner ◽  
...  
Keyword(s):  
Light Sheet ◽  
Fluorescence Excitation ◽  
Light Sheet Microscopy ◽  
Contrast Enhanced ◽  
Enhanced Imaging

scholarly journals Light sheet fluorescence microscopy of optically cleared brains for studying the glymphatic system

2020 ◽  
Vol 40 (10) ◽  
pp. 1975-1986
Author(s):  
Nicholas B Bèchet ◽  
Tekla M Kylkilahti ◽  
Bengt Mattsson ◽  
Martina Petrasova ◽  
Nagesh C Shanbhag ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Fluid Transport ◽  
Light Sheet ◽  
Brain Parenchyma ◽  
Sleep State ◽  
Glymphatic System ◽  
Light Sheet Microscopy ◽  
Highly Active ◽  
Quantitative Analyses ◽  
Light Sheet Fluorescence Microscopy

Fluid transport in the perivascular space by the glia-lymphatic (glymphatic) system is important for the removal of solutes from the brain parenchyma, including peptides such as amyloid-beta which are implicated in the pathogenesis of Alzheimer’s disease. The glymphatic system is highly active in the sleep state and under the influence of certain of anaesthetics, while it is suppressed in the awake state and by other anaesthetics. Here we investigated whether light sheet fluorescence microscopy of whole optically cleared murine brains was capable of detecting glymphatic differences in sleep- and awake-mimicking anaesthesia, respectively. Using light-sheet imaging of whole brains, we found anaesthetic-dependent cerebrospinal fluid (CSF) influx differences, including reduced tracer influx along tertiary branches of the middle cerebral artery and reduced influx along dorsal and anterior penetrating arterioles, in the awake-mimicking anaesthesia. This study establishes that light sheet microscopy of optically cleared brains is feasible for quantitative analyses and can provide images of the entire glymphatic system in whole brains.

scholarly journals Light-sheet microscopy in thick media using scanned Bessel beams and two-photon fluorescence excitation

2013 ◽  
Vol 21 (11) ◽  
pp. 13824 ◽  
Author(s):  
Florian O. Fahrbach ◽  
Vasily Gurchenkov ◽  
Kevin Alessandri ◽  
Pierre Nassoy ◽  
Alexander Rohrbach
Keyword(s):  
Light Sheet ◽  
Bessel Beams ◽  
Fluorescence Excitation ◽  
Two Photon ◽  
Light Sheet Microscopy ◽  
Two Photon Fluorescence ◽  
Photon Fluorescence

scholarly journals Wavefront shaping of a Bessel light field enhances light sheet microscopy with scattered light

2014 ◽  
Author(s):  
J. Nylk ◽  
C. Mitchell ◽  
T. Vettenburg ◽  
F. J. Gunn-Moore ◽  
K. Dholakia
Keyword(s):  
Light Field ◽  
Scattered Light ◽  
Light Sheet ◽  
Light Sheet Microscopy ◽  
Wavefront Shaping

scholarly journals Nonlinear light-sheet fluorescence microscopy by photobleaching imprinting

2014 ◽  
Vol 11 (93) ◽  
pp. 20130851 ◽  
Author(s):  
Liang Gao ◽  
Liren Zhu ◽  
Chiye Li ◽  
Lihong V. Wang
Keyword(s):  
Fluorescence Microscopy ◽  
Biological Samples ◽  
Scattered Light ◽  
Light Sheet ◽  
Fluorescence Decay ◽  
Field Of View ◽  
Optical Section ◽  
Ballistic Regime ◽  
Application Field ◽  
Light Sheet Fluorescence Microscopy

We present a nonlinear light-sheet fluorescence microscopy (LSFM) scheme based on photobleaching imprinting. By measuring photobleaching-induced fluorescence decay, our method simultaneously achieves a large imaging field of view and a thin optical section. Furthermore, the scattered-light-induced background is significantly reduced, considerably improving image contrast. Our method is expected to expand the application field of LSFM into the optical quasi-ballistic regime, enabling studies on non-transparent biological samples.

scholarly journals Light-Sheet Microscopy for Surface Topography Measurements and Quantitative Analysis

Sensors ◽  
2020 ◽  
Vol 20 (10) ◽  
pp. 2842 ◽  
Author(s):  
Zhanpeng Xu ◽  
Erik Forsberg ◽  
Yang Guo ◽  
Fuhong Cai ◽  
Sailing He
Keyword(s):  
Surface Topography ◽  
Spatial Resolution ◽  
Scattered Light ◽  
Three Dimensional ◽  
Light Sheet ◽  
Field Of View ◽  
Large Field ◽  
Linear Calibration ◽  
Industrial Sample ◽  
Light Sheet Microscopy

A novel light-sheet microscopy (LSM) system that uses the laser triangulation method to quantitatively calculate and analyze the surface topography of opaque samples is discussed. A spatial resolution of at least 10 μm in z-direction, 10 μm in x-direction and 25 μm in y-direction with a large field-of-view (FOV) is achieved. A set of sample measurements that verify the system′s functionality in various applications are presented. The system has a simple mechanical structure, such that the spatial resolution is easily improved by replacement of the objective, and a linear calibration formula, which enables convenient system calibration. As implemented, the system has strong potential for, e.g., industrial sample line inspections, however, since the method utilizes reflected/scattered light, it also has the potential for three-dimensional analysis of translucent and layered structures.

scholarly journals Enhanced Light Sheet Elastic Scattering Microscopy by Using a Supercontinuum Laser

2019 ◽  
Vol 2 (3) ◽  
pp. 57 ◽  
Author(s):  
Diego Di Battista ◽  
David Merino ◽  
Giannis Zacharakis ◽  
Pablo Loza-Alvarez ◽  
Omar E. Olarte
Keyword(s):  
Elastic Scattering ◽  
Spatial Scales ◽  
Scattered Light ◽  
Spatial Coherence ◽  
Light Sheet ◽  
Structural Features ◽  
Light Sheet Microscopy ◽  
Excitation Laser ◽  
Polarization Filtering ◽  
Microscopy Techniques

Light sheet fluorescence microscopy techniques have revolutionized biological microscopy enabling low-phototoxic long-term 3D imaging of living samples. Although there exist many light sheet microscopy (LSM) implementations relying on fluorescence, just a few works have paid attention to the laser elastic scattering source of contrast available in every light sheet microscope. Interestingly, elastic scattering can potentially disclose valuable information from the structure and composition of the sample at different spatial scales. However, when coherent scattered light is detected with a camera sensor, a speckled intensity is generated on top of the native imaged features, compromising their visibility. In this work, we propose a novel light sheet based optical setup which implements three strategies for dealing with speckles of elastic scattering images: (i) polarization filtering; (ii) reducing the temporal coherence of the excitation laser light; and, (iii) reducing the spatial coherence of the light sheet. Finally, we show how these strategies enable pristine light-sheet elastic-scattering imaging of structural features in challenging biological samples avoiding the deleterious effects of speckle, and without relying on, but complementing, fluorescent labelling.

scholarly journals Three-photon light-sheet fluorescence microscopy

2018 ◽  
Author(s):  
Adriá Escobet-Montalbán ◽  
Federico M. Gasparoli ◽  
Jonathan Nylk ◽  
Pengfei Liu ◽  
Zhengyi Yang ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Imaging Modality ◽  
Light Sheet ◽  
Wide Field ◽  
Excitation Light ◽  
Volumetric Imaging ◽  
Cellular Spheroids ◽  
Light Sheet Microscopy ◽  
Photon Excitation ◽  
Light Sheet Fluorescence Microscopy

We present the first demonstration of three-photon excitation light-sheet fluorescence microscopy. Light-sheet fluorescence microscopy in single- and two-photon modes has emerged as a powerful wide-field, low photo-damage technique for fast volumetric imaging of biological samples. We extend this imaging modality to the three-photon regime enhancing its penetration depth. Our present study uses a standard conventional femtosecond pulsed laser at 1000 nm wavelength for the imaging of 450 µm diameter cellular spheroids. In addition, we show, experimentally and through numerical simulations, the potential advantages in three-photon light-sheet microscopy of using propagation-invariant Bessel beams in preference to Gaussian beams.

scholarly journals Achromatic terahertz Airy beam generation with dielectric metasurfaces

Nanophotonics ◽  
2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Qingqing Cheng ◽  
Juncheng Wang ◽  
Ling Ma ◽  
Zhixiong Shen ◽  
Jing Zhang ◽  
...  
Keyword(s):  
Biomedical Imaging ◽  
Frequency Dispersion ◽  
Light Sheet ◽  
Photonic Devices ◽  
Self Healing ◽  
Beam Focusing ◽  
Light Sheet Microscopy ◽  
Airy Beam ◽  
Potential Applications ◽  
Airy Beams

AbstractAiry beams exhibit intriguing properties such as nonspreading, self-bending, and self-healing and have attracted considerable recent interest because of their many potential applications in photonics, such as to beam focusing, light-sheet microscopy, and biomedical imaging. However, previous approaches to generate Airy beams using photonic structures have suffered from severe chromatic problems arising from strong frequency dispersion of the scatterers. Here, we design and fabricate a metasurface composed of silicon posts for the frequency range 0.4–0.8 THz in transmission mode, and we experimentally demonstrate achromatic Airy beams exhibiting autofocusing properties. We further show numerically that a generated achromatic Airy-beam-based metalens exhibits self-healing properties that are immune to scattering by particles and that it also possesses a larger depth of focus than a traditional metalens. Our results pave the way to the realization of flat photonic devices for applications to noninvasive biomedical imaging and light-sheet microscopy, and we provide a numerical demonstration of a device protocol.

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