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Laser capture microscopy

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https://www.readbyqxmd.com/read/28501997/using-laser-capture-microdissection-to-isolate-cortical-laminae-in-nonhuman-primate-brain
#1
Brian A Corgiat, Claudius Mueller
Laser capture microdissection (LCM) is a technique that allows procurement of an enriched cell population from a heterogeneous tissue sample under direct microscopic visualization. Fundamentally, laser capture microdissection consists of three main steps: (1) visualizing the desired cell population by microscopy, (2) melting a thermolabile polymer onto the desired cell populations using infrared laser energy to form a polymer-cell composite (capture method) or photovolatizing a region of tissue using ultraviolet laser energy (cutting method), and (3) removing the desired cell population from the heterogeneous tissue...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28491859/autophagy-in-kidney-transplants-of-sirolimus-treated-recipients
#2
Sagar Bhayana, Arpita Baisantry, Thomas D Kraemer, Christoph Wrede, Jan Hegermann, Jan-Hinrich Bräsen, Clemens Bockmeyer, Jan Ulrich Becker, Matthias Ochs, Wilfried Gwinner, Hermann Haller, Anette Melk, Roland Schmitt
BACKGROUND: Mammalian target of rapamycin (mTOR) inhibitors are increasingly used as immunosuppressive agents in kidney transplantation. In the experimental setting it has been shown that mTOR inhibitors promote autophagy, but the concept that this might also occur in transplant patients has not been addressed. OBJECTIVES: This study was designed to investigate the association between mTOR inhibition and autophagy in renal transplants under routine clinical conditions...
March 2017: Journal of Nephropathology
https://www.readbyqxmd.com/read/28447987/live-cell-imaging-and-3d-analysis-of-angiotensin-receptor-type-1a-trafficking-in-transfected-human-embryonic-kidney-cells-using-confocal-microscopy
#3
Parnika Kadam, Ryan McAllister, Jeffrey S Urbach, Kathryn Sandberg, Susette C Mueller
Live-cell imaging is used to simultaneously capture time-lapse images of angiotensin type 1a receptors (AT1aR) and intracellular compartments in transfected human embryonic kidney-293 (HEK) cells following stimulation with angiotensin II (Ang II). HEK cells are transiently transfected with plasmid DNA containing AT1aR tagged with enhanced green fluorescent protein (EGFP). Lysosomes are identified with a red fluorescent dye. Live-cell images are captured on a laser scanning confocal microscope after Ang II stimulation and analyzed by software in three dimensions (3D, voxels) over time...
March 27, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28436646/filaments-with-affinity-binding-and-wet-strength-can-be-achieved-by-spinning-bifunctional-cellulose-nanofibrils
#4
Maija Vuoriluoto, Hannes Orelma, Meri Lundahl, Maryam Borghei, Orlando J Rojas
We demonstrate benzophenone (BP) conjugation via amine-reactive esters onto oxidized cellulosic fibers that were used as precursors, after microfluidization, of photoactive cellulose nanofibrils (CNF). From these fibrils, cellulose I filaments were synthesized by hydrogel spinning in an antisolvent followed by fast biradical UV cross-linking. As a result, the wet BP-CNF filaments retained extensively the original dry strength (a remarkable ∼80% retention). Thus, the principal limitation of these emerging materials was overcome (the wet tensile strength is typically <0...
May 3, 2017: Biomacromolecules
https://www.readbyqxmd.com/read/28399039/characterization-of-corneal-involvement-in-eyes-with-mucous-membrane-pemphigoid-by-in-vivo-confocal-microscopy
#5
Tudor C Tepelus, Jianyan Huang, SriniVas R Sadda, Olivia L Lee
PURPOSE: To describe the morphological features of the corneal epithelial layers, subbasal nerve plexus, stroma, and endothelium in patients with mucous membrane pemphigoid (MMP) as shown by in vivo confocal microscopy (IVCM). METHODS: Central corneal images were captured from 10 healthy age-matched control eyes and 30 eyes with clinically diagnosed MMP by in vivo laser scanning confocal microscopy (HRT III RCM). Morphological changes of the corneal epithelial layers, stroma, and endothelium, characteristics of corneal nerves, and presence of inflammatory dendritic cells (DCs) were evaluated...
April 10, 2017: Cornea
https://www.readbyqxmd.com/read/28395300/noninvasive-quantification-of-retinal-microglia-using-widefield-autofluorescence-imaging
#6
Despina Kokona, Nadia Schneider, Helena Giannakaki-Zimmermann, Joel Jovanovic, Andreas Ebneter, Martin Zinkernagel
Purpose: To validate widefield autofluorescence (AF) in vivo imaging of the retina in mice expressing green fluorescent protein (gfp) in microglia, and to monitor retinal microglia reconstitution in vivo after lethal irradiation and bone marrow transplantation. Methods: Transgenic Cx3cr1gfp/gfp and wildtype Balb/c mice were used in this study. A confocal scanning laser ophthalmoscope was used for AF imaging with a 55° and a widefield 102° lens. Intrasession reproducibility was assessed for each lens...
April 1, 2017: Investigative Ophthalmology & Visual Science
https://www.readbyqxmd.com/read/28364649/biochar-and-enhanced-phosphate-capture-mapping-mechanisms-to-functional-properties
#7
Jessica G Shepherd, Stephen Joseph, Saran P Sohi, Kate V Heal
A multi-technique analysis was performed on a range of biochar materials derived from secondary organic resources and aimed at sustainable recovery and re-use of wastewater phosphorus (P). Our purpose was to identify mechanisms of P capture in biochar and thereby inform its future optimisation as a sustainable P fertiliser. The biochar feedstock comprised pellets of anaerobically digested sewage sludge (PAD) or pellets of the same blended in the ratio 9:1 with ochre sourced from minewater treatment (POCAD), components which have limited alternative economic value...
July 2017: Chemosphere
https://www.readbyqxmd.com/read/28294302/polyclonal-and-monoclonal-igg-binding-on-protein-a-resins-evidence-of-competitive-binding-effects
#8
Justin Weinberg, Shaojie Zhang, Gillian Crews, Edward Healy, Giorgio Carta, Todd Przybycien
Protein A (ProA) chromatography is used extensively in the biopharmaceutical industry for the selective capture of both polyclonal and monoclonal antibodies (mAbs). This work provides a comparison of the adsorptive behavior of a highly heterogeneous polyclonal hIgG versus that of a mAb as well as the behavior of their mixtures on representative ProA resins. Both pH gradient elution and frontal loading experiments using human polyclonal IgG (hIgG) reveal a distribution of IgG-ProA binding strengths likely associated with multiple IgG subclasses and the heterogeneity of the variable region...
March 14, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28275314/ebm-regeneration-and-changes-in-ebm-component-mrna-expression-in-stromal-cells-after-corneal-injury
#9
Abirami Santhanam, Gustavo K Marino, Andre A M Torricelli, Steven E Wilson
PURPOSE: To investigate the production of the epithelial basement membrane (EBM) component mRNAs at time points before lamina lucida and lamina densa regeneration in anterior stromal cells after corneal injury that would heal with and without fibrosis. METHODS: Rabbit corneas were removed from 2 to 19 days after -4.5D or -9.0D photorefractive keratectomy (PRK) with the VISX S4 IR laser. Corneas were evaluated with transmission electron microscopy (TEM) for full regeneration of the lamina lucida and the lamina densa...
2017: Molecular Vision
https://www.readbyqxmd.com/read/28256794/laser-capture-microdissection-enables-transcriptomic-analysis-of-dividing-and-quiescent-liver-stages-of-plasmodium-relapsing-species
#10
Roger Cubi, Shruthi S Vembar, Anne Biton, Jean-Francois Franetich, Mallaury Bordessoulles, Daniel Sossau, Gigliola Zhanghi, Henriette Bosson-Vanga, Magalie Benard, Alicia Moreno, Nathalie Dereuddre-Bosquet, Roger Le Grand, Artur Scherf, Dominique Mazier
Dormant liver stage forms (hypnozoites) of the malaria parasite Plasmodium vivax present major hurdles to control and eradicate infection. Despite major research efforts, the molecular composition of hypnozoites remains ill defined. Here, we applied a combination of state-of-the-art technologies to generate the first transcriptome of hypnozoites. We developed a robust laser dissection microscopy protocol to isolate individual P. cynomolgi hypnozoites and schizonts from infected monkey hepatocytes and optimized RNA-seq analysis to obtain the first transcriptomes of these stages...
March 3, 2017: Cellular Microbiology
https://www.readbyqxmd.com/read/28250408/video-rate-volumetric-functional-imaging-of-the-brain-at-synaptic-resolution
#11
Rongwen Lu, Wenzhi Sun, Yajie Liang, Aaron Kerlin, Jens Bierfeld, Johannes D Seelig, Daniel E Wilson, Benjamin Scholl, Boaz Mohar, Masashi Tanimoto, Minoru Koyama, David Fitzpatrick, Michael B Orger, Na Ji
Neurons and neural networks often extend hundreds of micrometers in three dimensions. Capturing the calcium transients associated with their activity requires volume imaging methods with subsecond temporal resolution. Such speed is a challenge for conventional two-photon laser-scanning microscopy, because it depends on serial focal scanning in 3D and indicators with limited brightness. Here we present an optical module that is easily integrated into standard two-photon laser-scanning microscopes to generate an axially elongated Bessel focus, which when scanned in 2D turns frame rate into volume rate...
April 2017: Nature Neuroscience
https://www.readbyqxmd.com/read/28212742/demonstration-of-tissue-resident-memory-cd8-t-cells-in-insulitic-lesions-in-adult-patients-with-recent-onset-type-1-diabetes
#12
Enida Kuric, Peter Seiron, Lars Krogvold, Bjørn Edwin, Trond Buanes, Kristian F Hanssen, Oskar Skog, Knut Dahl-Jørgensen, Olle Korsgren
Subtypes of CD8(+) T cells in insulitic lesions in biopsy specimens from six subjects with recent-onset type 1 diabetes (T1D) and six nondiabetic matched controls were analyzed using simultaneous multicolor immunofluorescence. Also, insulitic islets based on accumulation of CD3(+) T cells were microdissected with laser-capture microscopy, and gene transcripts associated with inflammation and autoimmunity were analyzed. We found a substantial proportion, 43%, of the CD8(+) T cells in the insulitic lesions to display a tissue resident memory T cell (TRM) (CD8(+)CD69(+)CD103(+)) phenotype in T1D subjects...
March 2017: American Journal of Pathology
https://www.readbyqxmd.com/read/28196296/age-related-reduction-of-dermal-fibroblast-size-up-regulates-multiple-matrix-metalloproteinases-as-observed-in-aged-human-skin-in-vivo
#13
Z Qin, R M Balimunkwe, T Quan
BACKGROUND: Fragmentation of collagen fibrils, the major structure protein in skin, is a hallmark of dermal aging. Matrix metalloproteinases (MMPs) are largely responsible for fragmentation of collagen fibrils. However, the alteration of all known mammalian MMPs and the mechanism underlying altered expression of MMPs in chronologically aged human skin are less understood. OBJECTIVES: To quantify gene expression of all 23 known mammalian MMPs in sun-protected young and aged human skin in vivo, and investigate the potential mechanism underlying age-related alteration of multiple MMPs...
February 14, 2017: British Journal of Dermatology
https://www.readbyqxmd.com/read/28164461/flexible-polygon-mirror-based-laser-scanning-microscope-platform-for-multiphoton-in-vivo-imaging
#14
Y X Li, V Gautam, A Brüstle, I A Cockburn, V R Daria, C Gillespie, K Gaus, C Alt, W M Lee
Commercial microscopy systems make use of tandem scanning i.e. either slow or fast scanning. We constructed, for the first time, an advanced control system capable of delivering a dynamic line scanning speed ranging from 2.7 kHz to 27 kHz and achieve variable frame rates from 5 Hz to 50 Hz (512 × 512). The dynamic scanning ability is digitally controlled by a new customized open-source software named PScan1.0. This permits manipulation of scanning rates either to gain higher fluorescence signal at slow frame rate without increasing laser power or increase frame rates to capture high speed events...
February 6, 2017: Journal of Biophotonics
https://www.readbyqxmd.com/read/28107406/classification-and-segmentation-of-nanoparticle-diffusion-trajectories-in-cellular-micro-environments
#15
Thorsten Wagner, Alexandra Kroll, Chandrashekara R Haramagatti, Hans-Gerd Lipinski, Martin Wiemann
Darkfield and confocal laser scanning microscopy both allow for a simultaneous observation of live cells and single nanoparticles. Accordingly, a characterization of nanoparticle uptake and intracellular mobility appears possible within living cells. Single particle tracking allows to measure the size of a diffusing particle close to a cell. However, within the more complex system of a cell's cytoplasm normal, confined or anomalous diffusion together with directed motion may occur. In this work we present a method to automatically classify and segment single trajectories into their respective motion types...
2017: PloS One
https://www.readbyqxmd.com/read/28094895/a-simple-2d-composite-image-analysis-technique-for-the-crystal-growth-study-of-l-ascorbic-acid
#16
Krishan Kumar, Virender Kumar, Jatin Lal, Harmeet Kaur, Jasbir Singh
This work was destined for 2D crystal growth studies of l-ascorbic acid using the composite image analysis technique. Growth experiments on the l-ascorbic acid crystals were carried out by standard (optical) microscopy, laser diffraction analysis, and composite image analysis. For image analysis, the growth of l-ascorbic acid crystals was captured as digital 2D RGB images, which were then processed to composite images. After processing, the crystal boundaries emerged as white lines against the black (cancelled) background...
January 17, 2017: Microscopy Research and Technique
https://www.readbyqxmd.com/read/28068290/a-collection-system-for-dry-solid-residues-from-exhaled-breath-for-analysis-via-atomic-force-microscopy
#17
Victor N Morozov, Andrey Y Mikheev
Exhaled air contains sub-micron droplets of lung liquid, which potentially bear biomarkers of lung diseases. After dehydration they form dry residue particles (DRPs). As a first step in developing techniques to characterize individual DRPs, a new electrostatic collector was designed in which DRPs are charged within a unipolar corona charger, concentrated in a cone funnel, and deposited onto a limited area of a highly oriented pyrolytic graphite surface. The collector captures 80%-90% of DRPs at an optimal flow rate of 0...
January 9, 2017: Journal of Breath Research
https://www.readbyqxmd.com/read/28043041/detection-of-tissue-factor-positive-extracellular-vesicles-by-laser-scanning-confocal-microscopy
#18
Yohei Hisada, Alyson C Auriemma, Wyeth Alexander, Cihan Ay, Nigel Mackman
INTRODUCTION: Increased levels of tissue factor-positive extracellular vesicles (TF+EVs) have been detected in the plasma of patients with various diseases, including cancer and endotoxemia. Levels of TF+EVs in plasma samples can be measured by antigen and activity assays. The aim of the present study was to visualize TF+EVs by laser scanning confocal microscopy (LSCM). METHODS: EVs were isolated from the supernatant of two cultured human pancreatic cancer cell lines (Panc-1 and BxPc-3), from untreated or lipopolysaccharide (LPS) treated whole blood, and from plasma of pancreatic cancer patients...
February 2017: Thrombosis Research
https://www.readbyqxmd.com/read/27991699/a-supercritical-lens-optical-label-free-microscopy-sub-diffraction-resolution-and-ultra-long-working-distance
#19
Fei Qin, Kun Huang, Jianfeng Wu, Jinghua Teng, Cheng-Wei Qiu, Minghui Hong
A planar metalens for achieving super-resolution imaging in far-field is proposed. This metalens, which has a non-sub-wavelength feature size, can be fabricated by conventional laser pattern generator. The imaging process is purely physical and captured in real time, without any pre- and post-processing.
February 2017: Advanced Materials
https://www.readbyqxmd.com/read/27976852/four-dimensional-ultrafast-electron-microscopy-insights-into-an-emerging-technique
#20
COMMENT
Aniruddha Adhikari, Jeffrey K Eliason, Jingya Sun, Riya Bose, David J Flannigan, Omar F Mohammed
Four-dimensional ultrafast electron microscopy (4D-UEM) is a novel analytical technique that aims to fulfill the long-held dream of researchers to investigate materials at extremely short spatial and temporal resolutions by integrating the excellent spatial resolution of electron microscopes with the temporal resolution of ultrafast femtosecond laser-based spectroscopy. The ingenious use of pulsed photoelectrons to probe surfaces and volumes of materials enables time-resolved snapshots of the dynamics to be captured in a way hitherto impossible by other conventional techniques...
January 11, 2017: ACS Applied Materials & Interfaces
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