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Light sheet microscopy

Zer Vue, Gabriel Gonzalez, C Allison Stewart, Shyamin Mehra, Richard R Behringer
Endometrial or uterine glands secrete substances essential for uterine receptivity to the embryo, implantation, conceptus survival, and growth. Adenogenesis is the process of gland formation within the stroma of the uterus. In the mouse, uterine gland formation initiates at postnatal day (P) 5. Uterine gland morphology is poorly understood because it is primarily based on two-dimensional (2D) histology. To more fully describe uterine gland morphogenesis, we generated three-dimensional (3D) models of postnatal uterine glands from P0 to P21, based on volumetric imaging using light sheet microscopy...
March 15, 2018: Molecular Reproduction and Development
Paolo Baesso, Ricardo S Randall, Giovanni Sena
Light sheet fluorescence microscopy (LSFM) allows sustained and repeated optical sectioning of living specimens at high spatial and temporal resolution, with minimal photodamage. Here, we describe in detail both the hardware and the software elements of a live imaging method based on LSFM and optimized for tracking and 3D scanning of Arabidopsis root tips grown vertically in physiological conditions. The system is relatively inexpensive and with minimal footprint; hence it is well suited for laboratories of any size...
2018: Methods in Molecular Biology
Nadav Yayon, Amir Dudai, Nora Vrieler, Oren Amsalem, Michael London, Hermona Soreq
Three-dimensional structures in biological systems are routinely evaluated using large image stacks acquired from fluorescence microscopy; however, analysis of such data is muddled by variability in the signal across and between samples. Here, we present Intensify3D: a user-guided normalization algorithm tailored for overcoming common heterogeneities in large image stacks. We demonstrate the use of Intensify3D for analyzing cholinergic interneurons of adult murine brains in 2-Photon and Light-Sheet fluorescence microscopy, as well as of mammary gland and heart tissues...
March 9, 2018: Scientific Reports
C H Harrison, G R Buckland, S E Brooks, D A Johnston, D S Chatelet, A K L Liu, S M Gentleman, D Boche, J A R Nicoll
Current tissue-clearing protocols for imaging in three dimensions (3D) are typically applied to optimally fixed, small-volume rodent brain tissue - which is not representative of the tissue found in diagnostic neuropathology laboratories. We present a method to visualise the cerebral cortical vasculature in 3D in human post-mortem brain tissue which had been preserved in formalin for many years. Tissue blocks of cerebral cortex from two control cases, two Alzheimer's brains and two cases from Alzheimer's patients immunised against Aβ42 were stained with fluorescent Lycopersicon esculentum agglutinin (Tomato lectin), dehydrated and cleared using an adapted three-dimensional imaging of solvent cleared organs (3DISCO) protocol to visualise the vascular endothelium...
March 9, 2018: Journal of Anatomy
Nobuyuki Tanaka, Dagmara Kaczynska, Shigeaki Kanatani, Cecilia Sahlgren, Przemysław Mitura, Andrzej Stepulak, Ayako Miyakawa, Peter Wiklund, Per Uhlén
BACKGROUND: Cancers are heterogeneous and contain various types of irregular structures that can go undetected when examining them with standard two-dimensional microscopes. Studies of intricate networks of vasculature systems, e.g., the tumour lymphatic microvessels, benefit largely from three-dimensional imaging data analysis. METHODS: The new DIPCO (Diagnosing Immunolabeled Paraffin-Embedded Cleared Organs) imaging platform uses three-dimensional light-sheet microscopy and whole-mount immunolabelling of cleared samples to study proteins and micro-anatomies deep inside of tumours...
March 8, 2018: British Journal of Cancer
Mohammad Khursheed Siddiqi, Parvez Alam, Sumit Kumar Chaturvedi, Mohsin Vahid Khan, Saima Nusrat, Sadia Malik, Rizwan Hasan Khan
Protein aggregation and amyloid fibrillation are responsible for several serious pathological conditions (like type II diabetes, Alzheimer's and Parkinson's diseases etc.) and protein drugs ineffectiveness. Therefore, a molecule that can inhibit the amyloid fibrillation and potentially clear amyloid fibrils is of great therapeutic value. In this manuscript, we investigated the antiamyloidogenic, fibril disaggregating, as well as cell protective effect of an anti-tuberculosis drug, Capreomycin (CN). Aggregation kinetics data, as monitored by ThT fluorescence, inferred that CN retards the insulin amyloid fibrillation by primarily targeting the fibril elongation step with little effect on lag time...
February 26, 2018: Biochimica et Biophysica Acta
Tanner C Fadero, Therese M Gerbich, Kishan Rana, Aussie Suzuki, Matthew DiSalvo, Kristina N Schaefer, Jennifer K Heppert, Thomas C Boothby, Bob Goldstein, Mark Peifer, Nancy L Allbritton, Amy S Gladfelter, Amy S Maddox, Paul S Maddox
Fluorescence microscopy is a powerful approach for studying subcellular dynamics at high spatiotemporal resolution; however, conventional fluorescence microscopy techniques are light-intensive and introduce unnecessary photodamage. Light-sheet fluorescence microscopy (LSFM) mitigates these problems by selectively illuminating the focal plane of the detection objective by using orthogonal excitation. Orthogonal excitation requires geometries that physically limit the detection objective numerical aperture (NA), thereby limiting both light-gathering efficiency (brightness) and native spatial resolution...
February 28, 2018: Journal of Cell Biology
Kate McArthur, Lachlan W Whitehead, John M Heddleston, Lucy Li, Benjamin S Padman, Viola Oorschot, Niall D Geoghegan, Stephane Chappaz, Sophia Davidson, Hui San Chin, Rachael M Lane, Marija Dramicanin, Tahnee L Saunders, Canny Sugiana, Romina Lessene, Laura D Osellame, Teng-Leong Chew, Grant Dewson, Michael Lazarou, Georg Ramm, Guillaume Lessene, Michael T Ryan, Kelly L Rogers, Mark F van Delft, Benjamin T Kile
Mitochondrial apoptosis is mediated by BAK and BAX, two proteins that induce mitochondrial outer membrane permeabilization, leading to cytochrome c release and activation of apoptotic caspases. In the absence of active caspases, mitochondrial DNA (mtDNA) triggers the innate immune cGAS/STING pathway, causing dying cells to secrete type I interferon. How cGAS gains access to mtDNA remains unclear. We used live-cell lattice light-sheet microscopy to examine the mitochondrial network in mouse embryonic fibroblasts...
February 23, 2018: Science
Patrick Vilela, Amelie Heuer-Jungemann, Afaf El-Sagheer, Tom Brown, Otto L Muskens, Neil R Smyth, Antonios G Kanaras
Wound healing is a highly complex biological process, which is accompanied by changes in cell phenotype, variations in protein expression, and the production of active biomolecules. Currently, the detection of proteins in cells is done by immunostaining where the proteins in fixed cells are detected by labeled antibodies. However, immunostaining cannot provide information about dynamic processes in living cells, within the whole tissue. Here, an easy method is presented to detect the transition of epithelial to mesenchymal cells during wound healing...
February 21, 2018: Small
Marie-Pierre Adam, Marie Caroline Müllenbroich, Antonino Paolo Di Giovanna, Domenico Alfieri, Ludovico Silvestri, Leonardo Sacconi, Francesco Saverio Pavone
Although perfectly transparent specimens are imaged faster with light-sheet microscopy, less transparent samples are often imaged with two-photon microscopy leveraging its robustness to scattering; however, at the price of increased acquisition times. Clearing methods that are capable of rendering strongly scattering samples such as brain tissue perfectly transparent specimens are often complex, costly, and time intensive, even though for many applications a slightly lower level of tissue transparency is sufficient and easily achieved with simpler and faster methods...
February 2018: Journal of Biomedical Optics
A J P van Heugten, J Landman, A V Petukhov, H Vromans
The rheological properties of petrolatum are dependent on both temperature and thermal history. How this thermal dependency can be explained is unclear. In the past it has been suggested that the structure of petrolatum consists of a three-dimensional crystalline network. This has been established using old microscopic techniques only. Therefore a study on the microstructure of petrolatum was conducted using rheometry, DSC, pulsed NMR, polarized light microscopy and synchrotron X-ray. The combination of these techniques show that petrolatum is composed of 21% solid material at room temperature...
February 13, 2018: International Journal of Pharmaceutics
Jose M González-Domínguez, Verónica León, María Isabel Lucío, Maurizio Prato, Ester Vázquez
Graphene has promising physical and chemical properties such as high strength and flexibility, coupled with high electrical and thermal conductivities. It is therefore being incorporated into polymer-based composites for use in electronics and photonics applications. A main constraint related to the graphene development is that, being of a strongly hydrophobic nature, almost all dispersions (usually required for its handling and processing toward the desired application) are prepared in poisonous organic solvents such as N-methyl pyrrolidone or N,N-dimethyl formamide...
March 2018: Nature Protocols
Craig T Russell, Eric J Rees, Clemens F Kaminski
We compare the performance of linear and nonlinear methods for aligning the excitation and detection planes throughout volumes of large specimens in digitally scanned light sheet microscopy. An effective nonlinear method involves the registration of four corner extrema of the imaging volume via a projective transform. We show that this improves the light collection efficiency of the commonly used three-point affine registration by an average of 42% over a typical specimen volume. Accurate illumination/detection registration methods are now pertinent to biological research in view of current trends towards imaging large or expanded samples, at depth, with diffraction limited resolution...
February 15, 2018: Optics Letters
David Twapokera Mzinza, Henrike Fleige, Kristin Laarmann, Stefanie Willenzon, Jasmin Ristenpart, Julia Spanier, Gerd Sutter, Ulrich Kalinke, Peter Valentin-Weigand, Reinhold Förster
Bronchus-associated lymphoid tissue (BALT) develops at unpredictable locations around lung bronchi following pulmonary inflammation. The formation and composition of BALT have primarily been investigated by immunohistology that, due to the size of the invested organ, is usually restricted to a limited number of histological sections. To assess the entire BALT of the lung, other approaches are urgently needed. Here, we introduce a novel light sheet microscopy-based approach for assessing lymphoid tissue in the lung...
February 12, 2018: Cellular & Molecular Immunology
Mandana Ohadi, Gholamreza Dehghannoudeh, Hamid Forootanfar, Mojtaba Shakibaie, Majid Rajaee
In the present study the produced biosurfactant of Acinetobacter junii B6 (recently isolated from Iranian oil excavation site) were partially purified and identified by high performance thin layer chromatography (HPTLC), Fourier transform infrared spectroscopy (FTIR), and proton nuclear magnetic resonance (1H NMR). Elemental analysis of the biosurfactant by energy dispersive X-ray spectroscopy (EDS) revealed that the biosurfactant was anionic in nature. The physiochemical properties of the lipopeptide biosurfactant were evaluated by determination of its critical micelle concentration (CMC) and hydrophile-lipophile balance (HLB)...
February 6, 2018: International Journal of Biological Macromolecules
Sebastian J Streichan, Matthew Lefebvre, Nicholas Noll, Eric F Wieschaus, Boris I Shraiman
During embryogenesis tissue layers undergo morphogenetic flow rearranging and folding into specific shapes. While developmental biology has identified key genes and local cellular processes, global coordination of tissue remodeling at the organ scale remains unclear. Here we combine in toto light-sheet microscopy of the Drosophila embryo with quantitative analysis and physical modeling to relate cellular flow with the patterns of force generation during the gastrulation process. We find that the complex spatio-temporal flow pattern can be predicted from the measured meso-scale myosin density and anisotropy using a simple effective viscous model of the tissue, achieving close to 90% accuracy with one time dependent and two constant parameters...
February 9, 2018: ELife
Takayasu Kawasaki, Toyonari Yaji, Toshiaki Ohta, Koichi Tsukiyama, Kazuhiro Nakamura
Structure of amyloid β (Aβ) fibrils is rigidly stacked by β-sheet conformation, and the fibril state of Aβ is profoundly related to pathogenesis of Alzheimer's disease (AD). Although mid-infrared light has been used for various biological researches, it has not yet been known whether the infrared light changes the fibril structure of Aβ. In this study, we tested the effect of irradiation of intense mid-infrared light from a free-electron laser (FEL) targeting the amide bond on the reduction of β-sheet content in Aβ fibrils...
February 5, 2018: Cellular and Molecular Neurobiology
Khaled Khairy, William Lemon, Fernando Amat, Philipp J Keller
Mechanics plays a key role in the development of higher organisms. However, understanding this relationship is complicated by the difficulty of modeling the link between local forces generated at the subcellular level and deformations observed at the tissue and whole-embryo levels. Here we propose an approach first developed for lipid bilayers and cell membranes, in which force-generation by cytoskeletal elements enters a continuum mechanics formulation for the full system in the form of local changes in preferred curvature...
January 23, 2018: Biophysical Journal
Gregory Farber, Romulo Hurtado, Sarah Loh, Sébastien Monette, James Mtui, Raphael Kopan, Susan Quaggin, Catherine Meyer-Schwesinger, Doris Herzlinger, Rizaldy P Scott, Carl P Blobel
The principal function of glomeruli is to filter blood through a highly specialized filtration barrier consisting of a fenestrated endothelium, the glomerular basement membrane and podocyte foot processes. Previous studies have uncovered a crucial role of endothelial a disintegrin and metalloprotease 10 (ADAM10) and Notch signaling in the development of glomeruli, yet the resulting defects have not been further characterized nor understood in the context of kidney development. Here, we used several different experimental approaches to analyze the kidneys and glomeruli from mice lacking ADAM10 in endothelial cells (A10ΔEC mice)...
February 3, 2018: Angiogenesis
S Abadie, C Jardet, J Colombelli, B Chaput, A David, J-L Grolleau, P Bedos, V Lobjois, P Descargues, J Rouquette
BACKGROUND: Human skin is composed of the superimposition of tissue layers of various thicknesses and components. Histological staining of skin sections is the benchmark approach to analyse the organization and integrity of human skin biopsies; however, this approach does not allow 3D tissue visualization. Alternatively, confocal or two-photon microscopy is an effective approach to perform fluorescent-based 3D imaging. However, owing to light scattering, these methods display limited light penetration in depth...
January 27, 2018: Skin Research and Technology
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