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Light sheet microscopy

Duncan P Ryan, Elizabeth A Gould, Gregory J Seedorf, Omid Masihzadeh, Steven H Abman, Sukumar Vijayaraghavan, Wendy B Macklin, Diego Restrepo, Douglas P Shepherd
Optical tissue clearing has revolutionized researchers' ability to perform fluorescent measurements of molecules, cells, and structures within intact tissue. One common complication to all optically cleared tissue is a spatially heterogeneous refractive index, leading to light scattering and first-order defocus. We designed C-DSLM (cleared tissue digital scanned light-sheet microscopy) as a low-cost method intended to automatically generate in-focus images of cleared tissue. We demonstrate the flexibility and power of C-DSLM by quantifying fluorescent features in tissue from multiple animal models using refractive index matched and mismatched microscope objectives...
September 20, 2017: Nature Communications
Zhuang-Li Kang, Xiang Li, Hong-Ju He, Han-Jun Ma, Zhao-Jun Song
A comprehensive study was conducted to evaluate the structural changes of meat and protein of pork batters produced by chopping or beating process through the phase-contrast micrograph, laser light scattering analyzer, scanning electronic microscopy and Raman spectrometer. The results showed that the shattered myofibrilla fragments were shorter and particle-sizes were smaller in the raw batter produced by beating process than those in the chopping process. Compared with the raw and cooked batters produced by chopping process, modifications in amide I and amide III bands revealed a significant decrease of α-helix content and an increase of β-sheet, β-turn and random coils content in the beating process...
August 2017: Journal of Food Science and Technology
Shan-Shan Zhou, Shou-Qing Liu
Cocatalysts are vital for improving photocatalytic activity. Incorporating nitrogen atoms on a graphene frame using an imidazole cycle resulted in a new N-doped graphene (denoted as ING). Cerium(iv) oxide (CeO2) nanoparticles were dispersed on ING sheets, producing an ING/CeO2 hybrid material. The ING/CeO2 hybrid material was characterized using X-ray diffraction, transmission electron microscopy, Raman spectroscopy, ultraviolet-visible diffuse reflectance spectroscopy and surface photovoltage spectroscopy...
September 8, 2017: Photochemical & Photobiological Sciences
Collin Kieffer, Mark S Ladinsky, Pamela J Bjorkman
HIV transmission within lymphoid tissues remains incompletely characterized at the level of individual cells and virions. Here we visually describe our approach to understanding HIV-1 dissemination at different levels of volume and resolution within lymphoid tissues from HIV-1-infected humanized mice. We combined tissue clearing techniques, immunostaining, and light sheet fluorescence microscopy to visualize large-volumes of intact tissue with single-cell resolution from HIV-1-infected humanized mice. In parallel, we imaged adjacent regions of tissue using electron microscopy and electron tomography to gain 3D ultrastructural information about the same tissue samples...
September 7, 2017: AIDS Research and Human Retroviruses
Xinbo Zhuang, Wangang Zhang, Rui Liu, Yafu Liu, Lujuan Xing, Minyi Han, Zhuang-Li Kang, Xing-Lian Xu, Guang-Hong Zhou
The effects of sugarcane dietary fiber (SDF) on the gelation properties of porcine myofibrillar proteins (MP) were studied to understand its mechanism of action in improving gel functionality. Rheological tests on all composite gels (MP with SDF) showed the visco-elastic nature of MP, but the G' significantly increased with contents of SDF and with particle size (P<0.05). However, the δ exhibited the opposite effect. Light microscopy suggested that SDF affected moisture distribution in the gel by drawing water from MP and homogenously embedded in gelation...
October 2017: Food Research International
Sebastian Zundler, Anika Klingberg, Daniela Schillinger, Sarah Fischer, Clemens Neufert, Imke Atreya, Matthias Gunzer, Markus F Neurath
No abstract text is available yet for this article.
September 4, 2017: Gastroenterology
Elijah Shelton, Friedhelm Serwane, Otger Campàs
Modern fluorescence microscopy enables fast 3D imaging of biological and inert systems alike. In many studies, it is important to detect the surface of objects and quantitatively characterize its local geometry, including its mean curvature. We present a fully automated algorithm to determine the location and curvatures of an object from 3D fluorescence images, such as those obtained using confocal or light-sheet microscopy. The algorithm aims at reconstructing surface labelled objects with spherical topology and mild deformations from the spherical geometry with high accuracy, rather than reconstructing arbitrarily deformed objects with lower fidelity...
September 1, 2017: Journal of Microscopy
Søren S L Andersen
How axons are wired in the vertebrate spinal cord has been studied mostly using fixed samples or looking at individually growing axons. Using light-sheet microscopy, early neural development is here followed in vivo in real time at high resolution along several hundred micrometers of the zebrafish spinal cord. The dynamics and time course of cellular development and axonal wiring of interneurons expressing GFP under control of the dmrt3 promotor are analyzed. Following neurulation, commissural axons are observed crossing the ventral floor plate midline perpendicularly at about 20 microns/h and in a manner dependent on the Robo3 but not the EphA4 receptors...
August 30, 2017: Journal of Comparative Neurology
Annarita Falanga, Salvatore Valiante, Emilia Galdiero, Gianluigi Franci, Olga Scudiero, Giancarlo Morelli, Stefania Galdiero
gH625 constitutes a promising delivery vehicle for the transport of therapeutic biomacromolecules across membrane barriers. We report an application of multivalency to create a complex nanosystem for delivery and to elucidate the mechanism of peptide-lipid bilayer interactions. Multivalency may offer a route to enhance gH625 cellular uptake as demonstrated by results obtained on dimers of gH625 by fluorescence spectroscopy, circular dichroism, and surface plasmon resonance. Moreover, using both phase contrast and light sheet fluorescence microscopy we were able to characterize and visualize for the first time the fusion of giant unilamellar vesicles caused by a membranotropic peptide...
August 25, 2017: Scientific Reports
Rolf Fickentscher, Matthias Weiss
Asymmetric cell divisions are of fundamental importance for the development of multicellular organisms, e.g. for the generation of founder cells. Prime examples are asymmetric cell divisions in germline precursors during the early embryogenesis of the transparent roundworm Caenorhabditis elegans, one of the major developmental model organisms. However, due to a lack of quantitative data it has remained unclear how frequent unequal daughter cell sizes emerge in the worm's early embryogenesis, and whether these originate from sterical or biochemical cues...
August 24, 2017: Scientific Reports
Kevin M Dean, Reto Fiolka
We introduce a concept that enables parallelized three-dimensional imaging throughout large volumes with isotropic 300-350 nm resolution. By staggering high aspect ratio illumination beams laterally and axially within the depth of focus of a digitally scanned light-sheet fluorescence microscope (LSFM), multiple image planes can be simultaneously imaged with minimal cross-talk and light loss. We present a first demonstration of this concept for parallelized imaging by synthesizing two light-sheets with nonlinear Bessel beams and perform volumetric imaging of fluorescent beads and invasive breast cancer cells...
August 24, 2017: Scientific Reports
Natalia V Barykina, Oksana M Subach, Kiryl D Piatkevich, Erica E Jung, Aleksey Y Malyshev, Ivan V Smirnov, Andrey O Bogorodskiy, Valentin I Borshchevskiy, Anna M Varizhuk, Galina E Pozmogova, Edward S Boyden, Konstantin V Anokhin, Grigori N Enikolopov, Fedor V Subach
Currently available genetically encoded calcium indicators (GECIs) utilize calmodulins (CaMs) or troponin C from metazoa such as mammals, birds, and teleosts, as calcium-binding domains. The amino acid sequences of the metazoan calcium-binding domains are highly conserved, which may limit the range of the GECI key parameters and cause undesired interactions with the intracellular environment in mammalian cells. Here we have used fungi, evolutionary distinct organisms, to derive CaM and its binding partner domains and design new GECI with improved properties...
2017: PloS One
Michael J Taormina, Edouard A Hay, Raghuveer Parthasarathy
The fluids of the intestine serve as a physical barrier to pathogens, a medium for the diffusion of nutrients and metabolites, and an environment for commensal microbes. The rheological properties of intestinal mucus have therefore been the subject of many investigations, thus far limited to in vitro studies due to the difficulty of measurement in the natural context of the gut. This limitation especially hinders our understanding of how the gut microbiota interact with the intestinal space, since examination of this calls not only for in vivo measurement techniques, but for techniques that can be applied to model organisms in which the microbial state of the gut can be controlled...
August 22, 2017: Biophysical Journal
Mads Lichtenberg, Erik Christian Løvbjerg Trampe, Thomas Craig Vogelmann, Michael Kühl
In vivo variable-chlorophyll-fluorescence measurements of PSII quantum yields in optically dense systems are complicated by steep tissue light gradients due to scattering and absorption. Consequently, externally measured effective PSII quantum yields may be composed of signals derived from cells differentially exposed to actinic light, where cells located deeper inside tissues receive lower irradiance than cells closer to the surface, and can display distinct photophysiological status. We demonstrate how measured distributions of PSII quantum yields in plant tissue change under natural tissue light gradients as compared to conventionally measured quantum yields with even exposure to actinic light...
August 18, 2017: Plant Physiology
Serena Duchi, Filippo Piccinini, Michela Pierini, Alessandro Bevilacqua, Maria Luisa Torre, Enrico Lucarelli, Spartaco Santi
Cell interaction with biomaterials is one of the keystones to developing medical devices for tissue engineering applications. Biomaterials are the scaffolds that give three-dimensional support to the cells, and are vectors that deliver the cells to the injured tissue requiring repair. Features of biomaterials can influence the behaviour of the cells and consequently the efficacy of the tissue-engineered product. The adhesion, distribution and motility of the seeded cells onto the scaffold represent key aspects, and must be evaluated in vitro during the product development, especially when the efficacy of a specific tissue-engineered product depends on viable and functional cell loading...
2017: PloS One
René Hägerling, Dominik Drees, Aaron Scherzinger, Cathrin Dierkes, Silvia Martin-Almedina, Stefan Butz, Kristiana Gordon, Michael Schäfers, Klaus Hinrichs, Pia Ostergaard, Dietmar Vestweber, Tobias Goerge, Sahar Mansour, Xiaoyi Jiang, Peter S Mortimer, Friedemann Kiefer
BACKGROUND: Lack of investigatory and diagnostic tools has been a major contributing factor to the failure to mechanistically understand lymphedema and other lymphatic disorders in order to develop effective drug and surgical therapies. One difficulty has been understanding the true changes in lymph vessel pathology from standard 2D tissue sections. METHODS: VIPAR (volume information-based histopathological analysis by 3D reconstruction and data extraction), a light-sheet microscopy-based approach for the analysis of tissue biopsies, is based on digital reconstruction and visualization of microscopic image stacks...
August 17, 2017: JCI Insight
Sarah Schuster, Timothy Krüger, Ines Subota, Sina Thusek, Brice Rotureau, Andreas Beilhack, Markus Engstler
The highly motile and versatile protozoan pathogen Trypanosoma brucei undergoes a complex life cycle in the tsetse fly. Here we introduce the host insect as an expedient model environment for microswimmer research, as it allows examination of microbial motion within a diversified, secluded and yet microscopically tractable space. During their week-long journey through the different microenvironments of the fly´s interior organs, the incessantly swimming trypanosomes cross various barriers and confined surroundings, with concurrently occurring major changes of parasite cell architecture...
August 15, 2017: ELife
Sidra Islam, Moinuddin, Abdul Rouf Mir, Alok Raghav, Safia Habib, Khursheed Alam, Asif Ali
Glycation and oxidation induce structural alterations in the proteins in an inter-dependent manner with consequent pathological implications. The published literature presents wide range of modifications in conformational characteristics of proteins by glycation and oxidation; however, there is little data that could elaborate the cumulative effect of both the processes. This study has analysed the modifications in IgG by methylglyoxal (MG) (glycative stress), hydroxyl radical (OH(•)) (oxidative stress) and by their combined action i...
August 9, 2017: Journal of Biomolecular Structure & Dynamics
Deirdre Healy, Maria Nash, Alexander Gorleov, Kerry Thompson, Peter Dockery, Yury Rochev
The primary aim of this investigation was to determine the biocompatibility and cell culture potential of a newly designed class of thermoresponsive polymers. The attractiveness of these polymers lies in the fact that they swell rather than dissolve when the temperature is reduced below their respective lower critical solution temperature, due to the incorporation of octadecyl methacrylate (ODMA). The ODMA monomer acts as a physical crosslinker, preventing polymer dissolution upon temperature reduction. Two polymers were studied in this investigation poly(N isorpoylacrylamide (NIPAm)(99...
July 27, 2017: Colloids and Surfaces. B, Biointerfaces
Vipin Kumar, Liang Liu, Viet Cuong Nguyen, Venkateswarlu Bhavanasi, Kaushik Parida, Daniel Mandler, Pooi See Lee
Molybdenum trioxide is an interesting inorganic system in which the empty 4d states have potential to hold extra electrons and therefore can change states from insulating opaque (MoO3) to colored semimetallic (HxMoO3). Here, we characterize the local electrogeneration and charge transfer of the synthetic layered two-dimensional 2D MoO3-II (a polymorph of MoO3 and analogous to α-MoO3) in response to two different redox couples, i.e., [Ru(NH3)6](3+) and [Fe(CN)6](3-) by scanning electrochemical microscopy (SECM)...
August 16, 2017: ACS Applied Materials & Interfaces
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