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Intravital microscopy

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https://www.readbyqxmd.com/read/28723288/science-to-practice-can-mr-imaging-cell-tracking-of-macrophage-infiltration-be-used-as-a-predictive-imaging-biomarker-for-transplanted-stem-cell-rejection
#1
Jeff W M Bulte
Ferumoxytol-enhanced magnetic resonance (MR) imaging of donor-matched and mismatched stem cell transplants demonstrated decreased signal intensity not only for a xenogeneic mismatch in species but, surprisingly, also for a syngeneic mismatch in sex. MR imaging findings were corroborated with intravital fluorescence microscopy (IVM), where nearly 90% of all ferumoxytol-containing cells were found to be macrophages. Hence, MR imaging cell tracking of infiltrating macrophages may have predictive value in determining whether transplanted stem cell rejection will occur...
August 2017: Radiology
https://www.readbyqxmd.com/read/28720647/in-situ-targeting-of-dendritic-cells-sets-tolerogenic-environment-and-ameliorates-cd4-t-cell-response-in-the-postischemic-liver
#2
Dominik Funken, Hellen Ishikawa-Ankerhold, Bernd Uhl, Maximilian Lerchenberger, Markus Rentsch, Doris Mayr, Steffen Massberg, Jens Werner, Andrej Khandoga
CD4(+) T cells recruited to the liver play a key role in the pathogenesis of ischemia/reperfusion (I/R) injury. The mechanism of their activation during alloantigen-independent I/R is not completely understood. We hypothesized that liver-resident dendritic cells (DCs) interact with CD4(+) T cells in the postischemic liver and that modulation of DCs or T-cell-DC interactions attenuates liver inflammation. In mice, warm hepatic I/R (90/120-240 min) was induced. Tolerogenic DCs were generated in situ by pretreatment of animals with the vitamin D analog paricalcitol...
July 18, 2017: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
https://www.readbyqxmd.com/read/28720149/heparanase-promotes-neuroinflammatory-response-during-subarachnoid-hemorrhage-in-rats
#3
Benjarat Changyaleket, Zhao Zhong Chong, Randal O Dull, Danop Nanegrungsunk, Haoliang Xu
BACKGROUND: Heparanase, a mammalian endo-β-D-glucoronidase that specifically degrades heparan sulfate, has been implicated in inflammation and ischemic stroke. However, the role of heparanase in neuroinflammatory response in subarachnoid hemorrhage (SAH) has not yet been investigated. This study was designed to examine the association between heparanase expression and neuroinflammation during subarachnoid hemorrhage. METHODS: Rats were subjected to SAH by endovascular perforation, and the expression of heparanase was determined by Western blot analysis and immunofluorescence in the ipsilateral brain cortex at 24 h post-SAH...
July 18, 2017: Journal of Neuroinflammation
https://www.readbyqxmd.com/read/28715512/mouse-models-of-deep-vein-thrombosis
#4
T Schönfelder, S Jäckel, P Wenzel
The pathogenesis of venous thromboembolism (VTE) is still not completely understood. Experimental animals in which human deep vein thrombosis can be modeled are useful tools to investigate the pathogenesis of VTE. Besides the availability of transgenic and genetically modified mice, the use of high frequency ultrasound and intravital microscopy plays an important role in identifying thrombotic processes in mouse models. In this article, an overview about the application of various new technologies and existing mouse models is provided, and the impact of venous side branches on deep vein thrombosis in the mouse model is discussed...
2017: Gefässchirurgie: Zeitschrift Für Vaskuläre und Endovaskuläre Chirurgie
https://www.readbyqxmd.com/read/28699046/optical-clearing-and-fluorescence-deep-tissue-imaging-for-3d-quantitative-analysis-of-the-brain-tumor-microenvironment
#5
Tonny Lagerweij, Sophie A Dusoswa, Adrian Negrean, Esther M L Hendrikx, Helga E de Vries, Jeroen Kole, Juan J Garcia-Vallejo, Huibert D Mansvelder, W Peter Vandertop, David P Noske, Bakhos A Tannous, René J P Musters, Yvette van Kooyk, Pieter Wesseling, Xi Wen Zhao, Thomas Wurdinger
BACKGROUND: Three-dimensional visualization of the brain vasculature and its interactions with surrounding cells may shed light on diseases where aberrant microvascular organization is involved, including glioblastoma (GBM). Intravital confocal imaging allows 3D visualization of microvascular structures and migration of cells in the brain of mice, however, with limited imaging depth. To enable comprehensive analysis of GBM and the brain microenvironment, in-depth 3D imaging methods are needed...
July 11, 2017: Angiogenesis
https://www.readbyqxmd.com/read/28698070/surgical-preparation-of-rats-and-mice-for-intravital-microscopic-imaging-of-abdominal-organs
#6
George J Rhodes
Intravital microscopy is a powerful research tool that can provide insight into cellular and subcellular events that take place in organs in the body. However, meaningful results can only be obtained from animals whose physiology is preserved during the process of microscopy. Here I discuss the importance of preserving the overall state of health of the animal, methods of anesthesia, surgical techniques for intravital microscopy of various abdominal organs, methods to maintain and monitor the physiology of the animal during microscopy and associated peri- and post-operative recovery considerations...
July 8, 2017: Methods: a Companion to Methods in Enzymology
https://www.readbyqxmd.com/read/28691930/ly6clo-monocytes-drive-immunosuppression-and-confer-resistance-to-anti-vegfr2-cancer-therapy
#7
Keehoon Jung, Takahiro Heishi, Omar F Khan, Piotr S Kowalski, Joao Incio, Nuh N Rahbari, Euiheon Chung, Jeffrey W Clark, Christopher G Willett, Andrew D Luster, Seok Hyun Yun, Robert Langer, Daniel G Anderson, Timothy P Padera, Rakesh K Jain, Dai Fukumura
Current anti-VEGF therapies for colorectal cancer (CRC) provide limited survival benefit, as tumors rapidly develop resistance to these agents. Here, we have uncovered an immunosuppressive role for nonclassical Ly6Clo monocytes that mediates resistance to anti-VEGFR2 treatment. We found that the chemokine CX3CL1 was upregulated in both human and murine tumors following VEGF signaling blockade, resulting in recruitment of CX3CR1+Ly6Clo monocytes into the tumor. We also found that treatment with VEGFA reduced expression of CX3CL1 in endothelial cells in vitro...
July 10, 2017: Journal of Clinical Investigation
https://www.readbyqxmd.com/read/28691695/mechanical-and-biological-characterization-of-alkaline-substituted-orthophosphate-bone-substitutes-containing-meta-and-diphosphates
#8
Moritz Klein, Matthias W Laschke, Joerg H Holstein, Tina Histing, Tim Pohlemann, Michael Menger, Patric Garcia
Despite the growing knowledge on the mechanisms of fracture healing, bone defects often do not heal in a timely manner. Clinically, tricalcium phosphate (Ca-P) bone substitutes are used to fill bone defects and promote bone healing. However, the degradation rate of these implants is often too slow for sufficient bone replacement. The use of calcium phosphate material with the crystalline phase Ca10 [K/Na](PO4)7 containing different amounts of di- and metaphosphates may overcome this problem, because these materials show an accelerated degradation...
July 10, 2017: Biomedical Materials
https://www.readbyqxmd.com/read/28688659/indole-3-carbinol-is-a-potent-inhibitor-of-ischemia-reperfusion-induced-inflammation
#9
Emmanuel Ampofo, Nico Lachnitt, Jeannette Rudzitis-Auth, Beate M Schmitt, Michael D Menger, Matthias W Laschke
BACKGROUND: Ischemia-reperfusion (I/R) induces tissue inflammation, which is characterized by an increased leukocyte-endothelial cell interaction and leukocyte transmigration. These processes are mediated by the activation of the nuclear factor (NF)κB signaling pathway, resulting in an elevated expression of specific adhesion molecules. The phytochemical indole-3-carbinol (I3C) has been shown to exert anti-inflammatory effects by interfering with NFκB signal transduction. The aim of the present study was to investigate whether I3C is capable of counteracting the pathogenesis of I/R injury...
July 2017: Journal of Surgical Research
https://www.readbyqxmd.com/read/28683064/murine-chronic-lymph-node-window-for-longitudinal-intravital-lymph-node-imaging
#10
Eelco F J Meijer, Han-Sin Jeong, Ethel R Pereira, Thomas A Ruggieri, Cedric Blatter, Benjamin J Vakoc, Timothy P Padera
Chronic imaging windows in mice have been developed to allow intravital microscopy of many different organs and have proven to be of paramount importance in advancing our knowledge of normal and disease processes. A model system that allows long-term intravital imaging of lymph nodes would facilitate the study of cell behavior in lymph nodes during the generation of immune responses in a variety of disease settings and during the formation of metastatic lesions in cancer-bearing mice. We describe a chronic lymph node window (CLNW) surgical preparation that allows intravital imaging of the inguinal lymph node in mice...
August 2017: Nature Protocols
https://www.readbyqxmd.com/read/28671761/uptake-of-retinoic-acid-modified-pmma-nanoparticles-in-lx-2-and-liver-tissue-by-raman-imaging-and-intravital-microscopy
#11
Turgay Yildirim, Christian Matthäus, Adrian T Press, Stephanie Schubert, Michael Bauer, Jürgen Popp, Ulrich S Schubert
A primary amino-functionalized methyl methacrylate-based statistical copolymer is covalently coupled with retinoic acid (RA) and a fluorescent dye (DY590) in order to investigate the feasibility of the RA containing polymeric nanoparticles for Raman imaging studies and to study the possible selectivity of RA for hepatic stellate cells via intravital microscopy. Cationic nanoparticles are prepared by utilizing the nanoprecipitation method using modified polymers. Raman studies show that RA functional nanoparticles can be detectable in all tested cells without any need of additional label...
July 3, 2017: Macromolecular Bioscience
https://www.readbyqxmd.com/read/28669866/procedures-and-applications-of-long-term-intravital-microscopy
#12
REVIEW
Chloé Prunier, Na Chen, Laila Ritsma, Nienke Vrisekoop
Intravital microscopy (IVM) is increasingly used in biomedical research to study dynamic processes at cellular and subcellular resolution in their natural environment. Long-term IVM especially can be applied to visualize migration and proliferation over days to months within the same animal without recurrent surgeries. Skin can be repetitively imaged without surgery. To intermittently visualize cells in other organs, such as liver, mammary gland and brain, different imaging windows including the abdominal imaging window (AIW), dermal imaging window (DIW) and cranial imaging window (CIW) have been developed...
June 29, 2017: Methods: a Companion to Methods in Enzymology
https://www.readbyqxmd.com/read/28664407/combined-use-of-electron-microscopy-and-intravital-imaging-captures-morphological-and-functional-features-of-podocyte-detachment
#13
REVIEW
James L Burford, Georgina Gyarmati, Isao Shirato, Wilhelm Kriz, Kevin V Lemley, János Peti-Peterdi
The development of podocyte injury and albuminuria in various glomerular pathologies is still incompletely understood due to technical limitations in studying the glomerular filtration barrier (GFB) in real-time. We aimed to directly visualize the early morphological and functional changes of the GFB during the development of focal segmental glomerulosclerosis (FSGS) using a combination of transmission electron microscopy (TEM) and in vivo multiphoton microscopy (MPM) in the rat puromycin aminonucleoside (PAN) model...
June 29, 2017: Pflügers Archiv: European Journal of Physiology
https://www.readbyqxmd.com/read/28663906/fiber-bundle-based-integrated-platform-for-wide-field-fluorescence-imaging-and-patterned-optical-stimulation-for-modulation-of-vasoconstriction-in-the-deep-brain-of-a-living-animal
#14
Minkyung Kim, Jinki Hong, Jinsik Kim, Hyun-Joon Shin
We report a fiber optics-based intravital fluorescence imaging platform that includes epi-fluorescence microscopy and laser patterned-light stimulation system. The platform can perform real-time fluorescence imaging with a lateral resolution of ~4.9 μm while directly inserted into the intact mouse brain, optically stimulate vasoconstriction during real-time imaging, and avoid vessel damage in the penetration path of imaging probe. Using 473-nm patterned-light stimulation, we successfully modulated the vasoconstriction of a single targeted 37-μm-diameter blood vessel located more than 4...
June 1, 2017: Biomedical Optics Express
https://www.readbyqxmd.com/read/28661586/in-vivo-optical-imaging-intravital-microscopy
#15
EDITORIAL
Laura Marcu, Elizabeth M C Hillman
No abstract text is available yet for this article.
June 2017: Journal of Biophotonics
https://www.readbyqxmd.com/read/28626833/the-lung-is-a-host-defense-niche-for-immediate-neutrophil-mediated-vascular-protection
#16
Bryan G Yipp, Jung Hwan Kim, Ronald Lima, Lori D Zbytnuik, Björn Petri, Nick Swanlund, May Ho, Vivian G Szeto, Tamar Tak, Leo Koenderman, Peter Pickkers, Anton T J Tool, Taco W Kuijpers, Timo K van den Berg, Mark R Looney, Matthew F Krummel, Paul Kubes
Bloodstream infection is a hallmark of sepsis, a medically emergent condition requiring rapid treatment. However, upregulation of host defense proteins through toll-like receptors and NFκB requires hours after endotoxin detection. Using confocal pulmonary intravital microscopy, we identified that the lung provides a TLR4-Myd88-and abl tyrosine kinase-dependent niche for immediate CD11b-dependent neutrophil responses to endotoxin and Gram-negative bloodstream pathogens. In an in vivo model of bacteremia, neutrophils crawled to and rapidly phagocytosed Escherichia coli sequestered to the lung endothelium...
April 28, 2017: Science Immunology
https://www.readbyqxmd.com/read/28605364/quantitative-visualization-of-leukocyte-infiltrate-in-a-murine-model-of-fulminant-myocarditis-by-light-sheet-microscopy
#17
Linda Männ, Anika Klingberg, Matthias Gunzer, Mike Hasenberg
Light-sheet fluorescence microscopy (LSFM), in combination with chemical clearing protocols, has become the gold standard for analyzing fluorescently labelled structures in large biological specimens, and is down to cellular resolution. Meanwhile, the constant refinement of underlying protocols and the enhanced availability of specialized commercial systems enable us to investigate the microstructure of whole mouse organs and even allow for the characterization of cellular behavior in various live-cell imaging approaches...
May 31, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28600434/concerted-actions-of-distinct-nonmuscle-myosin-ii-isoforms-drive-intracellular-membrane-remodeling-in-live-animals
#18
Oleg Milberg, Akiko Shitara, Seham Ebrahim, Andrius Masedunskas, Muhibullah Tora, Duy T Tran, Yun Chen, Mary Anne Conti, Robert S Adelstein, Kelly G Ten Hagen, Roberto Weigert
Membrane remodeling plays a fundamental role during a variety of biological events. However, the dynamics and the molecular mechanisms regulating this process within cells in mammalian tissues in situ remain largely unknown. In this study, we use intravital subcellular microscopy in live mice to study the role of the actomyosin cytoskeleton in driving the remodeling of membranes of large secretory granules, which are integrated into the plasma membrane during regulated exocytosis. We show that two isoforms of nonmuscle myosin II, NMIIA and NMIIB, control distinct steps of the integration process...
July 3, 2017: Journal of Cell Biology
https://www.readbyqxmd.com/read/28589348/intravital-microscopy-of-t-b-cell-interactions-in-germinal-centers
#19
Changming Shih, Hai Qi
Dynamic imaging analyses of antigen-specific T-B interactions in germinal centers have advanced our understanding of the molecular mechanisms underlying affinity maturation and provided a wealth of information about how follicular helper T cells function in vivo. Here we describe a routine method to visualize fluorescence protein-expressing, antigen-specific T and B cells in germinal centers. The protocol for incorporating functional reporters or genetic perturbation of the T cells by retroviral transduction is also briefly described, using the FRET-based calcium reporter as an example...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28589347/fate-mapping-and-transcript-profiling-of-germinal-center-cells-by-two-photon-photoconversion
#20
Imogen Moran, Tri Giang Phan
The germinal center (GC) reaction is the key process for the generation of high affinity antibodies to foreign antigen. Standard experimental techniques such as fluorescence-activated cell sorting and histology have provided numerous insights into the composition and function of the GC. However, these approaches are limited to a "snapshot" in time and are unable to fully capture the dynamic nature of the GC. Intravital two-photon microscopy overcomes these disadvantages and has led to several major advances in the field but is restricted by practical and technical limits that prevent long-range mapping and molecular studies...
2017: Methods in Molecular Biology
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