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Laser capture microscopy

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https://www.readbyqxmd.com/read/29335532/intelligent-image-based-in-situ-single-cell-isolation
#1
Csilla Brasko, Kevin Smith, Csaba Molnar, Nora Farago, Lili Hegedus, Arpad Balind, Tamas Balassa, Abel Szkalisity, Farkas Sukosd, Katalin Kocsis, Balazs Balint, Lassi Paavolainen, Marton Z Enyedi, Istvan Nagy, Laszlo G Puskas, Lajos Haracska, Gabor Tamas, Peter Horvath
Quantifying heterogeneities within cell populations is important for many fields including cancer research and neurobiology; however, techniques to isolate individual cells are limited. Here, we describe a high-throughput, non-disruptive, and cost-effective isolation method that is capable of capturing individually targeted cells using widely available techniques. Using high-resolution microscopy, laser microcapture microscopy, image analysis, and machine learning, our technology enables scalable molecular genetic analysis of single cells, targetable by morphology or location within the sample...
January 15, 2018: Nature Communications
https://www.readbyqxmd.com/read/29328886/osteochondral-graft-size-is-significantly-associated-with-increased-force-and-decreased-chondrocyte-viability
#2
Brian E Walczak, Matthew S Nies, Darrin J Trask, Scott Hetzel, Patrick J Roney, Matthew W Squire, Geoffrey S Baer
BACKGROUND: Insertion force has been shown to significantly reduce chondrocyte viability during osteochondral allograft transplantation. How graft size influences the required insertion force and chondrocyte viability has yet to be determined. Hypothesis/Purpose: The purpose was to characterize how graft size influences insertion force requirements and chondrocyte viability during osteochondral transplantation. The hypothesis was that larger grafts would require greater force and reduce chondrocyte viability...
January 1, 2018: American Journal of Sports Medicine
https://www.readbyqxmd.com/read/29304390/the-cutting-edge-micro-ct-for-quantitative-toolmark-analysis-of-sharp-force-trauma-to-bone
#3
D G Norman, D G Watson, B Burnett, P M Fenne, M A Williams
Toolmark analysis involves examining marks created on an object to identify the likely tool responsible for creating those marks (e.g., a knife). Although a potentially powerful forensic tool, knife mark analysis is still in its infancy and the validation of imaging techniques as well as quantitative approaches is ongoing. This study builds on previous work by simulating real-world stabbings experimentally and statistically exploring quantitative toolmark properties, such as cut mark angle captured by micro-CT imaging, to predict the knife responsible...
December 30, 2017: Forensic Science International
https://www.readbyqxmd.com/read/29296484/fast-label-free-microscopy-technique-for-3d-dynamic-quantitative-imaging-of-living-cells
#4
José A Rodrigo, Juan M Soto, Tatiana Alieva
The refractive index (RI) is an important optical characteristic that is often exploited in label-free microscopy for analysis of biological objects. A technique for 3D RI reconstruction of living cells has to be fast enough to capture the cell dynamics and preferably needs to be compatible with standard wide-field microscopes. To solve this challenging problem, we present a technique that provides fast measurement and processing of data required for real-time 3D visualization of the object RI. Specifically, the 3D RI is reconstructed from the measurement of bright-field intensity images, axially scanned by a high-speed focus tunable lens mounted in front of a sCMOS camera, by using a direct deconvolution approach designed for partially coherent light microscopy in the non-paraxial regime...
December 1, 2017: Biomedical Optics Express
https://www.readbyqxmd.com/read/29259271/clearance-of-a-persistent-picornavirus-infection-is-associated-with-enhanced-pro-apoptotic-and-cellular-immune-responses
#5
Carolina Stenfeldt, Michael Eschbaumer, George R Smoliga, Luis L Rodriguez, James Zhu, Jonathan Arzt
Long-term persistent viral infections cause substantial morbidity and associated economic losses in human and veterinary contexts. Yet, the mechanisms associated with establishment of persistent infections are poorly elucidated. We investigated immunomodulatory mechanisms associated with clearance versus persistence of foot-and-mouth disease virus (FMDV) in micro-dissected compartments of the bovine nasopharynx by microarray. The use of laser-capture microdissection allowed elucidation of differential gene regulation within distinct anatomic compartments critical to FMDV infection...
December 19, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29258460/a-case-of-proliferative-diabetic-retinopathy-in-which-scintillating-particles-appeared-in-the-intravitreal-cavity-after-laser-photocoagulation
#6
Ryohsuke Kohmoto, Takatoshi Kobayashi, Takaki Sato, Daisaku Kimura, Masanori Fukumoto, Kensuke Tajiri, Teruyo Kida, Tsunehiko Ikeda
BACKGROUND: To report a case of proliferative diabetic retinopathy (PDR) exhibiting the appearance of scintillating particles presumed to be crystallin inside the intravitreal cavity after laser photocoagulation. CASE PRESENTATION: A 56-year-old male patient presented at our outpatient clinic after becoming aware of decreased vision in his right eye. Ocular examination performed at the patient's initial visit revealed a massive preretinal macular hemorrhage due to PDR in his right eye...
December 19, 2017: BMC Ophthalmology
https://www.readbyqxmd.com/read/29226243/single-frame-3d-fluorescence-microscopy-with-ultraminiature-lensless-flatscope
#7
Jesse K Adams, Vivek Boominathan, Benjamin W Avants, Daniel G Vercosa, Fan Ye, Richard G Baraniuk, Jacob T Robinson, Ashok Veeraraghavan
Modern biology increasingly relies on fluorescence microscopy, which is driving demand for smaller, lighter, and cheaper microscopes. However, traditional microscope architectures suffer from a fundamental trade-off: As lenses become smaller, they must either collect less light or image a smaller field of view. To break this fundamental trade-off between device size and performance, we present a new concept for three-dimensional (3D) fluorescence imaging that replaces lenses with an optimized amplitude mask placed a few hundred micrometers above the sensor and an efficient algorithm that can convert a single frame of captured sensor data into high-resolution 3D images...
December 2017: Science Advances
https://www.readbyqxmd.com/read/29182594/pharmacokinetics-of-chlorin-e%C3%A2-cobalt-bis-dicarbollide-conjugate-in-balb-c-mice-with-engrafted-carcinoma
#8
Arthur B Volovetsky, Vladimir S Sukhov, Irina V Balalaeva, Varvara V Dudenkova, Natalia Yu Shilyagina, Аlexey V Feofanov, Anastasija V Efremenko, Mikhail A Grin, Andrey F Mironov, Igor B Sivaev, Vladimir I Bregadze, Anna V Maslennikova
The necessary precondition for efficient boron neutron capture therapy (BNCT) is control over the content of isotope 10B in the tumor and normal tissues. In the case of boron-containing porphyrins, the fluorescent part of molecule can be used for quantitative assessment of the boron content. Study Objective: We performed a study of the biodistribution of the chlorin e₆-Cobalt bis(dicarbollide) conjugate in carcinoma-bearing Balb/c mice using ex vivo fluorescence imaging, and developed a mathematical model describing boron accumulation and release based on the obtained experimental data...
November 28, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/29118098/x-ray-free-electron-laser-opportunities-for-drug-discovery
#9
REVIEW
Robert K Y Cheng, Rafael Abela, Michael Hennig
Past decades have shown the impact of structural information derived from complexes of drug candidates with their protein targets to facilitate the discovery of safe and effective medicines. Despite recent developments in single particle cryo-electron microscopy, X-ray crystallography has been the main method to derive structural information. The unique properties of X-ray free electron laser (XFEL) with unmet peak brilliance and beam focus allow X-ray diffraction data recording and successful structure determination from smaller and weaker diffracting crystals shortening timelines in crystal optimization...
November 8, 2017: Essays in Biochemistry
https://www.readbyqxmd.com/read/29103202/stain-free-histopathology-of-basal-cell-carcinoma-by-dual-vibration-resonance-frequency-cars-microscopy
#10
Norbert Kiss, Ádám Krolopp, Kende Lőrincz, András Bánvölgyi, Róbert Szipőcs, Norbert Wikonkál
Basal cell carcinoma (BCC) is the most common malignancy in Caucasians. Nonlinear microscopy has been previously utilized for the imaging of BCC, but the captured images do not correlate with H&E staining. Recently, Freudiger et al. introduced a novel method to visualize tissue morphology analogous to H&E staining, using coherent anti-Stokes Raman scattering (CARS) technique. In our present work, we introduce a novel algorithm to post-process images obtained from dual vibration resonance frequency (DVRF) CARS measurements to acquire high-quality pseudo H&E images of BCC samples...
November 4, 2017: Pathology Oncology Research: POR
https://www.readbyqxmd.com/read/29101151/laser-scanning-in-vivo-confocal-microscopy-ivcm-for-evaluating-human-corneal-sub-basal-nerve-plexus-parameters-protocol-for-a-systematic-review
#11
Manikkuwadura Eranda Harshan De Silva, Alexis Ceecee Zhang, Amalia Karahalios, Holly Rose Chinnery, Laura Elizabeth Downie
INTRODUCTION: Laser scanning in vivo confocal microscopy (IVCM) enables non-invasive, high-resolution imaging of the cornea. In recent years, there has been a vast increase in researchers using laser scanning IVCM to image and quantify corneal nerve parameters. However, a range of methodological approaches have been adopted. The primary aim of this systematic review is to critically appraise the reported method(s) of primary research studies that have used laser scanning IVCM to quantify corneal sub-basal nerve plexus (SBNP) parameters in humans, and to examine corneal nerve parameters in healthy individuals...
November 3, 2017: BMJ Open
https://www.readbyqxmd.com/read/29043677/using-optical-tweezers-combined-with-total-internal-reflection-microscopy-to-study-interactions-between-the-er-and-golgi-in-plant-cells
#12
Imogen Sparkes, Rhiannon R White, Benjamin Coles, Stanley W Botchway, Andy Ward
Optical tweezers have been used to trap and micromanipulate several biological specimens ranging from DNA, macromolecules, organelles to single celled organisms. Using a combination of the refraction and scattering of laser light from a focused laser beam, refractile objects are physically captured and can be moved within the surrounding media. The technique is routinely used to determine biophysical properties such as the forces exerted by motor proteins. Here, we describe how optical tweezers combined with total internal reflection fluorescence (TIRF) microscopy can be used to assess physical interactions between organelles, more specifically the ER and Golgi bodies in plant cells...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28884710/atmospheric-pulsed-laser-deposition-and-thermal-annealing-of-plasmonic-silver-nanoparticle-films
#13
T M Khan, M A Mujawar, K E Siewerska, A Pokle, T Donnelly, N McEvoy, G S Duesberg, J G Lunney
A new method for pulsed laser deposition of plasmonic silver nanoparticle (NP) films in flowing gas at atmospheric pressure is described. The ablation was done using an excimer laser at 248 nm. Fast optical imaging shows that the ablation plume is captured by the flowing gas, and is expected to form a NP aerosol, which is carried 5-20 mm to the substrate. The dependence of the deposition rate on laser fluence, gas flow velocity, and target-substrate distance was investigated using electron microscopy and absorption spectroscopy of the deposited films...
November 3, 2017: Nanotechnology
https://www.readbyqxmd.com/read/28860973/particle-tracking-facilitates-real-time-capable-motion-correction-in-2d-or-3d-two-photon-imaging-of-neuronal-activity
#14
Samira Aghayee, Daniel E Winkowski, Zachary Bowen, Erin E Marshall, Matt J Harrington, Patrick O Kanold, Wolfgang Losert
The application of 2-photon laser scanning microscopy (TPLSM) techniques to measure the dynamics of cellular calcium signals in populations of neurons is an extremely powerful technique for characterizing neural activity within the central nervous system. The use of TPLSM on awake and behaving subjects promises new insights into how neural circuit elements cooperatively interact to form sensory perceptions and generate behavior. A major challenge in imaging such preparations is unavoidable animal and tissue movement, which leads to shifts in the imaging location (jitter)...
2017: Frontiers in Neural Circuits
https://www.readbyqxmd.com/read/28851950/biomechanical-properties-of-predator-induced-body-armour-in-the-freshwater-crustacean-daphnia
#15
Sebastian Kruppert, Martin Horstmann, Linda C Weiss, Ulrich Witzel, Clemens F Schaber, Stanislav N Gorb, Ralph Tollrian
The freshwater crustacean Daphnia is known for its ability to develop inducible morphological defences that thwart predators. These defences are developed only in the presence of predators and are realized as morphological shape alterations e.g. 'neckteeth' in D. pulex and 'crests' in D. longicephala. Both are discussed to hamper capture, handling or consumption by interfering with the predator's prey capture devices. Additionally, D. pulex and some other daphniids were found to armour-up and develop structural alterations resulting in increased carapace stiffness...
August 29, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28844788/spatiotemporal-resolution-of-spinal-meningeal-and-parenchymal-inflammation-during-experimental-autoimmune-encephalomyelitis
#16
Bandana Shrestha, Xi Jiang, Shujun Ge, Debayon Paul, Peter Chianchiano, Joel S Pachter
Experimental autoimmune encephalomyelitis (EAE) induced by active immunization of C57BL/6 mice with peptide from myelin oligodendrocyte protein (MOG35-55), is a neuroinflammatory, demyelinating disease widely recognized as an animal model of multiple sclerosis (MS). Typically, EAE presents with an ascending course of paralysis, and inflammation that is predominantly localized to the spinal cord. Recent studies have further indicated that inflammation - in both MS and EAE - might initiate within the meninges and propagate from there to the underlying parenchyma...
August 24, 2017: Neurobiology of Disease
https://www.readbyqxmd.com/read/28836259/cartilage-matrix-remodelling-differs-by-disease-state-and-joint-type
#17
M-F Hsueh, V B Kraus, P Önnerfjord
Dramatic alterations in mechanical properties have been documented for osteoarthritic (OA) cartilage. However, the matrix composition underlying these changes has not been mapped and their aetiology is not entirely understood. We hypothesised that an understanding of the cartilage matrix heterogeneity could provide insights into the origin of these OA-related alterations. We generated serial transverse cryo sections for 7 different cartilage conditions: 2 joint sites (knee and hip), 2 disease states (healthy and OA) and 3 tissue depths (superficial, middle and deep)...
August 24, 2017: European Cells & Materials
https://www.readbyqxmd.com/read/28829423/multi-photon-time-lapse-imaging-to-visualize-development-in-real-time-visualization-of-migrating-neural-crest-cells-in-zebrafish-embryos
#18
Antionette L Williams, Brenda L Bohnsack
Congenital eye and craniofacial anomalies reflect disruptions in the neural crest, a transient population of migratory stem cells that give rise to numerous cell types throughout the body. Understanding the biology of the neural crest has been limited, reflecting a lack of genetically tractable models that can be studied in vivo and in real-time. Zebrafish is a particularly important developmental model for studying migratory cell populations, such as the neural crest. To examine neural crest migration into the developing eye, a combination of the advanced optical techniques of laser scanning microscopy with long wavelength multi-photon fluorescence excitation was implemented to capture high-resolution, three-dimensional, real-time videos of the developing eye in transgenic zebrafish embryos, namely Tg(sox10:EGFP) and Tg(foxd3:GFP), as sox10 and foxd3 have been shown in numerous animal models to regulate early neural crest differentiation and likely represent markers for neural crest cells...
August 9, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28725020/neuronal-imaging-with-ultrahigh-dynamic-range-multiphoton-microscopy
#19
Ruohui Yang, Timothy D Weber, Ellen D Witkowski, Ian G Davison, Jerome Mertz
Multiphoton microscopes are hampered by limited dynamic range, preventing weak sample features from being detected in the presence of strong features, or preventing the capture of unpredictable bursts in sample strength. We present a digital electronic add-on technique that vastly improves the dynamic range of a multiphoton microscope while limiting potential photodamage. The add-on provides real-time negative feedback to regulate the laser power delivered to the sample, and a log representation of the sample strength to accommodate ultrahigh dynamic range without loss of information...
July 19, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28651457/metallosis-in-a-dog-as-a-long-term-complication-following-total-hip-arthroplasty
#20
Mallory J DiVincenzo, Galit H Frydman, Michael P Kowaleski, Charles R Vanderburg, Barry Lai, Trisha J Oura, Samuel H Jennings
Metallosis is the accumulation of metallic debris in soft tissues resulting from wear following total joint replacement. A dog was evaluated for lameness 4 years after total hip arthroplasty using a titanium alloy and cobalt chromium total hip system. Radiographs revealed severe acetabular component wear, implant-bone interface deterioration, and peri-acetabular osteolysis. During surgical revision, black periarticular tissue surrounded the implants. Histologically, there was fibrosis and granulomatous inflammation with abundant, intra- and extracellular, black, granular material and smaller amounts of clear punctate to acicular material...
January 1, 2017: Veterinary Pathology
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