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Light sheet microscopy

Jonas Doerr, Martin Karl Schwarz, Dirk Wiedermann, Anke Leinhaas, Alina Jakobs, Florian Schloen, Inna Schwarz, Michael Diedenhofen, Nils Christian Braun, Philipp Koch, Daniel A Peterson, Ulrich Kubitscheck, Mathias Hoehn, Oliver Brüstle
While transplantation represents a key tool for assessing in vivo functionality of neural stem cells and their suitability for neural repair, little is known about the integration of grafted neurons into the host brain circuitry. Rabies virus-based retrograde tracing has developed into a powerful approach for visualizing synaptically connected neurons. Here, we combine this technique with light sheet fluorescence microscopy (LSFM) to visualize transplanted cells and connected host neurons in whole-mouse brain preparations...
January 19, 2017: Nature Communications
Tobias Meinert, Benjamin Alexander Gutwein, Alexander Rohrbach
Light-sheet microscopy enables fast 3D, high-contrast imaging in biology and colloidal sciences. Recently, the controlled transport of living embryos or small colloids through stable glass capillaries is manifold interesting. Although they hardly impair the sample, glass capillaries spoil the image by generating significant aberrations of the illumination and detection light. Here, we analyze the deflection of illuminating Bessel beams at the capillary by k-spectral shifting, and correct for it by a beam deflector...
January 15, 2017: Optics Letters
Anand P Singh, Rémi Galland, Megan L Finch-Edmondson, Gianluca Grenci, Jean-Baptiste Sibarita, Vincent Studer, Virgile Viasnoff, Timothy E Saunders
The three-dimensional (3D) architecture of the cell nucleus plays an important role in protein dynamics and in regulating gene expression. However, protein dynamics within the 3D nucleus are poorly understood. Here, we present, to our knowledge, a novel combination of 1) single-objective based light-sheet microscopy, 2) photoconvertible proteins, and 3) fluorescence correlation microscopy, to quantitatively measure 3D protein dynamics in the nucleus. We are able to acquire >3400 autocorrelation functions at multiple spatial positions within a nucleus, without significant photobleaching, allowing us to make reliable estimates of diffusion dynamics...
January 10, 2017: Biophysical Journal
Emilio J Gualda, Hugo Pereira, Gabriel G Martins, Rui Gardner, Nuno Moreno
Flow cytometry is the tool of choice for high-speed acquisition and analysis of large cell populations, with the tradeoff of lacking intracellular spatial information. Although in the last decades flow cytometry systems that can actually acquire two-dimensional spatial information were developed, some of the limitations remained though, namely constrains related to sample size and lack of depth or dynamic information. The combination of fluidics and light-sheet illumination has the potential to address these limitations...
January 11, 2017: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
Manuel Schmitz, Ben K A Nelemans, Theodoor H Smit
Due to its availability, low cost, flat geometry, and transparency, the ex ovo chick embryo has become a major vertebrate animal model for the study of morphogenetic events, such as gastrulation(2), neurulation(3)(-)(5), somitogenesis(6), heart bending(7,8), and brain formation(9)(-)(13), during early embryogenesis. Key to understanding morphogenetic processes is to follow them dynamically by time-lapse imaging. The acquisition of time-lapse movies of chick embryogenesis ex ovo has been limited either to short time windows or to the need for an incubator to control temperature and humidity around the embryo(14)...
December 28, 2016: Journal of Visualized Experiments: JoVE
P Bruza, H Lin, L Jarvis, D Gladstone, B Pogue
PURPOSE: To recover a three-dimensional density distribution of luminescent molecular probes located several centimeters deep within a highly scattering tissue. METHODS: We developed a novel sheet beam Cherenkov-excited luminescence scanned imaging (CELSI) methodology. The sample was irradiated by a horizontally oriented, vertically scanned 6 MV X-ray sheet beam (200mm × 5mm, 0.2mm vertical step) from a radiotherapy linear accelerator. The resulting Cherenkov light emission - and thus luminescent probe excitation - occurred exclusively along the irradiation plane due to a short diffusion path of secondary particles and Cherenkov photons...
June 2016: Medical Physics
Pei Ma, Dennis C Chan, Shi Gu, Michiko Watanabe, Michael W Jenkins, Andrew M Rollins
Optical mapping (OM) of electrical activity using voltage-sensitive fluorescent dyes is a powerful tool for the investigation of embryonic cardiac electrophysiology. However, because conventional OM integrates the signal in depth and projects it to a two-dimensional plane, information acquired is incomplete and dependent upon the orientation of the sample. This complicates interpretation of data, especially when comparing one heart to another. To overcome this limitation, we present volumetric OM using light-sheet microscopy, which enables high-speed capture of optically sectioned slices...
December 1, 2016: Biomedical Optics Express
Vincent Rouger, Ricardo Alchini, Alexei Kazarine, Angelica A Gopal, Marie-Pier Girouard, Alyson E Fournier, Paul W Wiseman
No abstract text is available yet for this article.
December 1, 2016: Journal of Biomedical Optics
Jung-Hong Min, Hoe-Min Kwak, Kiyoung Kim, Woo-Lim Jeong, Dong-Seon Lee
In this paper, we introduce very thin Indium tin oxide (ITO) layers (5, 10, and 15 nm) hybridized with a metal mesh to produce high-performance transparent conductive layers (TCLs) in near-ultraviolet light-emitting diodes (NUV LEDs). Using UV-vis-IR spectrometry, Hall measurement, and atomic force microscopy, we found that 10 nm was the optimal thickness for the very thin ITO layers in terms of outstanding transmittance and sheet resistance values as well as stable contact properties when hybridized with the metal mesh...
January 27, 2017: Nanotechnology
Bethan Lloyd-Lewis, Felicity M Davis, Olivia B Harris, Jessica R Hitchcock, Filipe C Lourenco, Mathias Pasche, Christine J Watson
BACKGROUND: High-resolution 3D imaging of intact tissue facilitates cellular and subcellular analyses of complex structures within their native environment. However, difficulties associated with immunolabelling and imaging fluorescent proteins deep within whole organs have restricted their applications to thin sections or processed tissue preparations, precluding comprehensive and rapid 3D visualisation. Several tissue clearing methods have been established to circumvent issues associated with depth of imaging in opaque specimens...
December 13, 2016: Breast Cancer Research: BCR
Benjamin Wipfler, Hans Pohl, Margarita I Yavorskaya, Rolf G Beutel
Techniques currently used in insect morphology are outlined briefly. Scanning electron microscopy (SEM) and microphotography are used mainly for documenting external features, the former providing more information on tiny surface structures and the latter on coloration, transparency and degree of sclerotization. A broad spectrum of methods is now available for anatomical studies: histological serial sections, confocal laser scanning microscopy (CLSM), light-sheet fluorescence microscopy (LSFM), serial block-face scanning electron microscopy (SBFSEM), dual beam scanning electron microscopy (FIB-SEM), nuclear magnetic resonance imaging (NMRI), and μ-computed tomography (micro-CT)...
December 2016: Current Opinion in Insect Science
Frederic Strobl, Ernst Hk Stelzer
Light sheet-based fluorescence microscopy became an important tool in insect developmental biology due to its high acquisition speed, low photo-bleaching rate and the high survival probability of the specimens. Initially applied to document the embryogenesis of Drosophila melanogaster, it is now used to investigate the embryonic morphogenesis of emerging model organisms such as the red flour beetle Tribolium castaneum. Here, we discuss the principles of light sheet-based fluorescence microscopy and outline Tribolium as a model organism for developmental biology...
December 2016: Current Opinion in Insect Science
Louise Helene Søgaard Jensen, Lars Michael Skjolding, Amalie Thit, Sara Nørgaard Sørensen, Carsten Købler, Kristian Mølhave, Anders Baun
Increasing use of engineered nanoparticles has led to extensive research into their potential hazards to the environment and human health. Cellular uptake from the gut is sparsely investigated, and microscopy techniques applied for uptake studies can result in misinterpretations. Various microscopy techniques were used to investigate internalization of 10-nm gold nanoparticles in Daphnia magna gut lumen and gut epithelial cells following 24-h exposure and outline potential artifacts (i.e., high-contrast precipitates from sample preparation related to these techniques)...
November 25, 2016: Environmental Toxicology and Chemistry
Vincent Maioli, George Chennell, Hugh Sparks, Tobia Lana, Sunil Kumar, David Carling, Alessandro Sardini, Chris Dunsby
Light sheet fluorescence microscopy has previously been demonstrated on a commercially available inverted fluorescence microscope frame using the method of oblique plane microscopy (OPM). In this paper, OPM is adapted to allow time-lapse 3-D imaging of 3-D biological cultures in commercially available glass-bottomed 96-well plates using a stage-scanning OPM approach (ssOPM). Time-lapse 3-D imaging of multicellular spheroids expressing a glucose Förster resonance energy transfer (FRET) biosensor is demonstrated in 16 fields of view with image acquisition at 10 minute intervals...
November 25, 2016: Scientific Reports
Brianna M Harvey, Kunwar P Bhatnagar, Robert J Schenck, Alfred L Rosenberger, Susan J Rehorek, Anne M Burrows, Valerie B DeLeon, Timothy D Smith
Living primates have relatively large eyes and support orbital tissues with a postorbital bar (POB) and/or septum. Some mammals with large eyes lack a POB, and presumably rely on soft tissues. Here, we examined the orbits of four species of strepsirrhine primates (Galagidae, Cheirogaleidae) and three species of fruit bats (Pteropodidae). Microdissection and light microscopy were employed to identify support structures of the orbit. In bats and primates, there are two layers of fascial sheets that border the eye laterally...
December 2016: Anatomical Record: Advances in Integrative Anatomy and Evolutionary Biology
Juneyong Eum, Jina Kwak, Hee Joung Kim, Seoyoung Ki, Kooyeon Lee, Ahmed A Raslan, Ok Kyu Park, Md Ashraf Uddin Chowdhury, Song Her, Yun Kee, Seung-Hae Kwon, Byung Joon Hwang
Environmental contamination by trinitrotoluene is of global concern due to its widespread use in military ordnance and commercial explosives. Despite known long-term persistence in groundwater and soil, the toxicological profile of trinitrotoluene and other explosive wastes have not been systematically measured using in vivo biological assays. Zebrafish embryos are ideal model vertebrates for high-throughput toxicity screening and live in vivo imaging due to their small size and transparency during embryogenesis...
November 17, 2016: International Journal of Molecular Sciences
Jonathan Nylk, Kaley McCluskey, Sanya Aggarwal, Javier A Tello, Kishan Dholakia
We have investigated the effect of Airy illumination on the image quality and depth penetration of digitally scanned light-sheet microscopy in turbid neural tissue. We used Fourier analysis of images acquired using Gaussian and Airy light-sheets to assess their respective image quality versus penetration into the tissue. We observed a three-fold average improvement in image quality at 50 μm depth with the Airy light-sheet. We also used optical clearing to tune the scattering properties of the tissue and found that the improvement when using an Airy light-sheet is greater in the presence of stronger sample-induced aberrations...
October 1, 2016: Biomedical Optics Express
Anna Russo, Carlo Diaferia, Sara La Manna, Cinzia Giannini, Teresa Sibillano, Antonella Accardo, Giancarlo Morelli, Ettore Novellino, Daniela Marasco
Nucleophosmin (NPM1) is a multifunctional protein involved in a variety of biological processes including the pathogenesis of several human malignancies and is the most frequently mutated gene in Acute Myeloid Leukemia (AML). To deepen the role of protein regions in its biological activities, lately we reported on the structural behavior of dissected C-terminal domain (CTD) helical fragments. Unexpectedly the H2 (residues 264-277) and H3 AML-mutated regions showed a remarkable tendency to form amyloid-like assemblies with fibrillar morphology and β-sheet structure that resulted as toxic when exposed to human neuroblastoma cells...
February 2017: Biochimica et Biophysica Acta
Carlo Diaferia, Eliana Gianolio, Antonella Accardo, Giancarlo Morelli
Telechelic PEG-polymers end-capped by diphenylalanine (FF) motives and containing a DOTA-Gd complex, bound on a lysine side chain at the centre of peptide moiety, are studied for their assembling properties and for the relaxometric behavior. The observed variations in terms of relaxivity are correlated to the assembling properties of the aggregates by using several techniques: fluorescence, Circular Dichroism (CD) and Fourier Transform Infrared (FTIR) for aggregation tendency and secondary structure determination; Dynamic Light Scattering (DLS) and Transmission Electron Microscopy (TEM) for morphological definition...
November 15, 2016: Journal of Peptide Science: An Official Publication of the European Peptide Society
Soumik Siddhanta, Santosh Kumar Paidi, Kathryn Bushley, Ram Prasad, Ishan Barman
Imaging tip growth in fungal hyphae is highly warranted to unravel the molecular mechanism of this extraordinarily precise and localized phenomenon. In situ probing of fungal cultures, however, have been challenging due to their inherent complexity and light penetration issues associated with conventional optical imaging. In this work, we report a label-free approach using a combination of light sheet microscopy and Raman spectroscopy to obtain concomitant morphological and biochemical information from the growing specimen...
November 15, 2016: Chemphyschem: a European Journal of Chemical Physics and Physical Chemistry
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