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Laser capture microscopy

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https://www.readbyqxmd.com/read/28294302/polyclonal-and-monoclonal-igg-binding-on-protein-a-resins-evidence-of-competitive-binding-effects
#1
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
#2
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
#3
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
#4
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...
February 27, 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
#5
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
#6
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
#7
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
#8
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
#9
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
#10
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
#11
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
#12
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
#13
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 3, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/27958348/laser-light-field-fusion-for-wide-field-lensfree-on-chip-phase-contrast-microscopy-of-nanoparticles
#14
Farnoud Kazemzadeh, Alexander Wong
Wide-field lensfree on-chip microscopy, which leverages holography principles to capture interferometric light-field encodings without lenses, is an emerging imaging modality with widespread interest given the large field-of-view compared to lens-based techniques. In this study, we introduce the idea of laser light-field fusion for lensfree on-chip phase contrast microscopy for detecting nanoparticles, where interferometric laser light-field encodings acquired using a lensfree, on-chip setup with laser pulsations at different wavelengths are fused to produce marker-free phase contrast images of particles at the nanometer scale...
December 13, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27942581/b-cell-repertoire-expansion-occurs-in-meningeal-ectopic-lymphoid-tissue
#15
Klaus Lehmann-Horn, Sheng-Zhi Wang, Sharon A Sagan, Scott S Zamvil, H-Christian von Büdingen
Ectopic lymphoid tissues (ELT) can be found in multiple sclerosis (MS) and other organ-specific inflammatory conditions. Whether ELT in the meninges of central nervous system (CNS) autoimmune disease exhibit local germinal center (GC) activity remains unknown. In an experimental autoimmune encephalomyelitis model of CNS autoimmunity, we found activation-induced cytidine deaminase, a GC-defining enzyme, in meningeal ELT (mELT) densely populated by B and T cells. To determine GC activity in mELT, we excised meningeal lymphoid aggregates using laser capture microscopy and evaluated B cell repertoires in mELT and secondary lymphoid organs by next-generation immune repertoire sequencing...
December 8, 2016: JCI Insight
https://www.readbyqxmd.com/read/27896016/wide-field-fourier-ptychographic-microscopy-using-laser-illumination-source
#16
Jaebum Chung, Hangwen Lu, Xiaoze Ou, Haojiang Zhou, Changhuei Yang
Fourier ptychographic (FP) microscopy is a coherent imaging method that can synthesize an image with a higher bandwidth using multiple low-bandwidth images captured at different spatial frequency regions. The method's demand for multiple images drives the need for a brighter illumination scheme and a high-frame-rate camera for a faster acquisition. We report the use of a guided laser beam as an illumination source for an FP microscope. It uses a mirror array and a 2-dimensional scanning Galvo mirror system to provide a sample with plane-wave illuminations at diverse incidence angles...
November 1, 2016: Biomedical Optics Express
https://www.readbyqxmd.com/read/27890026/electrostatic-capture-following-laser-microdissection-for-the-preparation-of-homogeneous-biological-specimens
#17
Dingrong Yi, Linghua Kong, Ranjith K Kankala, Zi Wang
There is an unmet need for researchers in life sciences and clinical pathology to obtain untainted target cells with very high accuracy, which are suitable for subsequent genome and protein analysis. In this paper, an electrostatic capture laser microdissection technology (ECM) is proposed and explained. Following microscopic identification and separation of target cells from the surrounding tissues using laser cutting, the ECM uses electrostatic forces to capture target cells. Validation experiments indicate that ECM can capture a wide assortment of contamination-free homogeneous samples, ranging from very tiny pieces of a few micrometers in diameter to large pieces with a surface area of over 40,000 µm2...
December 2016: Microscopy and Microanalysis
https://www.readbyqxmd.com/read/27788214/image-based-modeling-of-blood-flow-and-oxygen-transfer-in-feto-placental-capillaries
#18
Philip Pearce, Paul Brownbill, Jiří Janáček, Marie Jirkovská, Lucie Kubínová, Igor L Chernyavsky, Oliver E Jensen
During pregnancy, oxygen diffuses from maternal to fetal blood through villous trees in the placenta. In this paper, we simulate blood flow and oxygen transfer in feto-placental capillaries by converting three-dimensional representations of villous and capillary surfaces, reconstructed from confocal laser scanning microscopy, to finite-element meshes, and calculating values of vascular flow resistance and total oxygen transfer. The relationship between the total oxygen transfer rate and the pressure drop through the capillary is shown to be captured across a wide range of pressure drops by physical scaling laws and an upper bound on the oxygen transfer rate...
2016: PloS One
https://www.readbyqxmd.com/read/27763643/mitochondrial-angiotensin-receptors-in-dopaminergic-neurons-role-in-cell-protection-and-aging-related-vulnerability-to-neurodegeneration
#19
Rita Valenzuela, Maria A Costa-Besada, Javier Iglesias-Gonzalez, Emma Perez-Costas, Begoña Villar-Cheda, Pablo Garrido-Gil, Miguel Melendez-Ferro, Ramon Soto-Otero, Jose L Lanciego, Daniel Henrion, Rafael Franco, Jose L Labandeira-Garcia
The renin-angiotensin system (RAS) was initially considered as a circulating humoral system controlling blood pressure, being kidney the key control organ. In addition to the 'classical' humoral RAS, a second level in RAS, local or tissular RAS, has been identified in a variety of tissues, in which local RAS play a key role in degenerative and aging-related diseases. The local brain RAS plays a major role in brain function and neurodegeneration. It is normally assumed that the effects are mediated by the cell-surface-specific G-protein-coupled angiotensin type 1 and 2 receptors (AT1 and AT2)...
October 20, 2016: Cell Death & Disease
https://www.readbyqxmd.com/read/27723140/synthesis-of-fluorescent-gelators-and-direct-observation-of-gelation-with-a-fluorescence-microscope
#20
Kenji Hanabusa, Takuya Ueda, Shingo Takata, Masahiro Suzuki
Fluorescein-, benzothiazole-, quinoline-, stilbene-, and carbazole-containing fluorescent gelators have been synthesized by connecting gelation-driving segments, including l-isoleucine, l-valine, l-phenylalanine, l-leucine residue, cyclo(l-asparaginyl-l-phenylalanyl), and trans-(1R,2R)-diaminocyclohexane. The emission behaviors of the gelators were investigated, and their gelation abilities studied against 15 solvents. The minimum gel concentration, variable-temperature spectroscopy, transmission electron microscopy, scanning electron microscopy, fluorescence microscopy (FM), and confocal laser scanning microscopy (CLSM) were used to characterize gelation...
November 14, 2016: Chemistry: a European Journal
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