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Intravital microscopy

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https://www.readbyqxmd.com/read/28626833/the-lung-is-a-host-defense-niche-for-immediate-neutrophil-mediated-vascular-protection
#1
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
#2
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
#3
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...
June 9, 2017: Journal of Cell Biology
https://www.readbyqxmd.com/read/28589348/intravital-microscopy-of-t-b-cell-interactions-in-germinal-centers
#4
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
#5
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
https://www.readbyqxmd.com/read/28561381/estrogen-dependent-efficacy-of-limb-ischemic-preconditioning-in-female-rats
#6
Levente Pócs, Ágnes Janovszky, Dénes Garab, Gabriella Terhes, Imre Ocsovszki, József Kaszaki, Mihály Boros, József Piffkó, Andrea Szabó
Our aim was to examine the effects of ischemic preconditioning (IPC) on the local periosteal and systemic inflammatory consequences of hindlimb ischemia-reperfusion (IR) in Sprague-Dawley rats with chronic estrogen deficiency (13 weeks after ovariectomy, OVX) in the presence and absence of chronic 17beta-estradiol supplementation (E2, 20 µg kg(-1) , 5 days/week for 5 weeks); sham-operated (non-OVX) animals served as controls. As assessed by intravital fluorescence microscopy, rolling and the firm adhesion of polymorphonuclear neutrophil leukocytes (PMNs) gave similar results in the Sham + IR and OVX + IR groups in the tibial periosteal microcirculation during the 3-h reperfusion period after a 60-min tourniquet ischemia...
May 31, 2017: Journal of Orthopaedic Research: Official Publication of the Orthopaedic Research Society
https://www.readbyqxmd.com/read/28552620/in-vivo-imaging-reveals-existence-of-crypt-fission-and-fusion-in-adult-mouse-intestine
#7
Lotte Bruens, Saskia I J Ellenbroek, Jacco van Rheenen, Hugo J Snippert
The intestinal epithelium is a repetitive sheet of crypt and villus units with stem cells at the bottom of the crypts. During postnatal development, crypts multiply via fission, generating 2 daughter crypts from 1 parental crypt. In the adult intestine, crypt fission is observed at a low frequency. Using intravital microscopy in Lgr5(EGFP-Ires-CreERT2) mice, we monitored individual crypt dynamics over multiple days with single-cell resolution. We discovered the existence of crypt fusion, an almost exact reverse phenomenon of crypt fission, in which 2 crypts fuse into 1 daughter crypt...
May 25, 2017: Gastroenterology
https://www.readbyqxmd.com/read/28552391/where-are-we-the-anatomy-of-the-murine-cortical-meninges-revisited-for-intravital-imaging-immunology-and-clearance-of-waste-from-the-brain
#8
REVIEW
Jonathan A Coles, Elmarie Myburgh, James M Brewer, Paul G McMenamin
Rapid progress is being made in understanding the roles of the cerebral meninges in the maintenance of normal brain function, in immune surveillance, and as a site of disease. Most basic research on the meninges and the neural brain is now done on mice, major attractions being the availability of reporter mice with fluorescent cells, and of a huge range of antibodies useful for immunocytochemistry and the characterization of isolated cells. In addition, two-photon microscopy through the unperforated calvaria allows intravital imaging of the undisturbed meninges with sub-micron resolution...
May 25, 2017: Progress in Neurobiology
https://www.readbyqxmd.com/read/28539925/cxcl9-derived-peptides-differentially-inhibit-neutrophil-migration-in-vivo-through-interference-with-glycosaminoglycan-interactions
#9
Vincent Vanheule, Daiane Boff, Anneleen Mortier, Rik Janssens, Björn Petri, Elzbieta Kolaczkowska, Paul Kubes, Nele Berghmans, Sofie Struyf, Andreas J Kungl, Mauro Martins Teixeira, Flavio Almeida Amaral, Paul Proost
Several acute and chronic inflammatory diseases are driven by accumulation of activated leukocytes due to enhanced chemokine expression. In addition to specific G protein-coupled receptor-dependent signaling, chemokine-glycosaminoglycan (GAG) interactions are important for chemokine activity in vivo. Therefore, the GAG-chemokine interaction has been explored as target for inhibition of chemokine activity. It was demonstrated that CXCL9(74-103) binds with high affinity to GAGs, competed with active chemokines for GAG binding and thereby inhibited CXCL8- and monosodium urate (MSU) crystal-induced neutrophil migration to joints...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/28538086/sphingosine-1-phosphate-receptor-1-agonist-sew2871-causes-severe-cardiac-side-effects-and-does-not-improve-microvascular-barrier-breakdown-in-sepsis
#10
Sven Flemming, Natalie Burkard, Michael Meir, Martin Alexander Schick, Christoph-Thomas Germer, Nicolas Schlegel
BACKGROUND: Endothelial barrier dysfunction is a hallmark in the pathogenesis of sepsis. Sphingosine-1-phosphate (S1P) has been proposed to be critically involved in the maintenance of endothelial barrier function predominately by activating S1P receptor-1 (S1P1). Previous studies have shown that the specific S1P1 agonist SEW2871 improves endothelial barrier function under inflammatory conditions. However, the effectiveness of SEW2871 and potential side effects remained largely unexplored in a clinically relevant model of sepsis...
May 23, 2017: Shock
https://www.readbyqxmd.com/read/28528637/find-your-way-with-x-ray-using-microct-to-correlate-in-vivo-imaging-with-3d-electron-microscopy
#11
Matthia A Karreman, Bernhard Ruthensteiner, Luc Mercier, Nicole L Schieber, Gergely Solecki, Frank Winkler, Jacky G Goetz, Yannick Schwab
Combining in vivo imaging with electron microscopy (EM) uniquely allows monitoring rare and critical events in living tissue, followed by their high-resolution visualization in their native context. A major hurdle, however, is to keep track of the region of interest (ROI) when moving from intravital microscopy (IVM) to EM. Here, we present a workflow that relies on correlating IVM and microscopic X-ray computed tomography to predict the position of the ROI inside the EM-processed sample. The ROI can then be accurately and quickly targeted using ultramicrotomy and imaged using EM...
2017: Methods in Cell Biology
https://www.readbyqxmd.com/read/28527736/the-effects-of-non-invasive-radiofrequency-electric-field-hyperthermia-on-biotransport-and-biodistribution-of-fluorescent-60-fullerene-derivative-in-a-murine-orthotopic-model-of-breast-adenocarcinoma
#12
Norman A Lapin, Martyna Krzykawska-Serda, Sean Dilliard, Yuri Mackeyev, Maciej Serda, Lon J Wilson, Steven A Curley, Stuart J Corr
The aim of this study is to understand the combined and differential biokinetic effects of radiofrequency (RF) electric-field hyperthermia as an adjunctive therapy to [60]fullerene nanoparticle-based drug delivery systems in targeting the micro-vasculature and micro-environments of breast cancer tumors. Intravital microscopy (IVM) is an ideal tool to provide the spatial and temporal resolution needed for quantification in this investigation. The water-soluble and fluorescent [60]fullerene derivative (C60-serPF) was designed to be an amphiphilic nanostructure, which is able to cross several biological membranes and accumulate in tumor tissues by passing through abnormally leaky tumor blood vessels...
May 17, 2017: Journal of Controlled Release: Official Journal of the Controlled Release Society
https://www.readbyqxmd.com/read/28526762/exacerbation-of-thromboinflammation-by-hyperglycemia-precipitates-cerebral-infarct-growth-and-hemorrhagic-transformation
#13
Jean-Philippe Desilles, Varouna Syvannarath, Véronique Ollivier, Clément Journé, Sandrine Delbosc, Célina Ducroux, William Boisseau, Liliane Louedec, Lucas Di Meglio, Stéphane Loyau, Martine Jandrot-Perrus, Louis Potier, Jean-Baptiste Michel, Mikael Mazighi, Benoit Ho-Tin-Noé
BACKGROUND AND PURPOSE: Admission hyperglycemia is associated with a poor outcome in acute ischemic stroke. How hyperglycemia impacts the pathophysiology of acute ischemic stroke remains largely unknown. We investigated how preexisting hyperglycemia increases ischemia/reperfusion cerebral injury. METHODS: Normoglycemic and streptozotocin-treated hyperglycemic rats were subjected to transient middle cerebral artery occlusion. Infarct growth and brain perfusion were assessed by magnetic resonance imaging...
May 19, 2017: Stroke; a Journal of Cerebral Circulation
https://www.readbyqxmd.com/read/28526034/differential-darc-ackr1-expression-distinguishes-venular-from-non-venular-endothelial-cells-in-murine-tissues
#14
Aude Thiriot, Carolina Perdomo, Guiying Cheng, Igor Novitzky-Basso, Sara McArdle, Jamie K Kishimoto, Olga Barreiro, Irina Mazo, Robinson Triboulet, Klaus Ley, Antal Rot, Ulrich H von Andrian
BACKGROUND: Intravascular leukocyte recruitment in most vertebrate tissues is restricted to postcapillary and collecting venules, whereas capillaries and arterioles usually support little or no leukocyte adhesion. This segmental restriction is thought to be mediated by endothelial, rather than hemodynamic, differences. The underlying mechanisms are largely unknown, in part because effective tools to distinguish, isolate, and analyze venular endothelial cells (V-ECs) and non-venular endothelial cells (NV-ECs) have been unavailable...
May 19, 2017: BMC Biology
https://www.readbyqxmd.com/read/28522327/measurement-of-bacterial-capture-and-phagosome-maturation-of-kupffer-cells-by-intravital-microscopy
#15
Bas G J Surewaard, Paul Kubes
It is central to the field of bacterial pathogenesis to define how bacteria are killed by phagocytic cells. During phagocytosis, the microbe is localized to the phagolysosome where crucial defense mechanisms such as acidification and production of reactive oxygen species (ROS) are initiated. This process has extensively been studied in vitro, however many resident tissue phagocytes will phenotypically change upon isolation from their natural environment. Therefore, interrogation of phagocytosis and phagosomal function of cells in the context of their natural tissue environment enhances our understanding of the biological process in vivo...
May 15, 2017: Methods: a Companion to Methods in Enzymology
https://www.readbyqxmd.com/read/28504989/structural-behavior-of-the-endothelial-glycocalyx-is-associated-with-pathophysiologic-status-in-septic-mice-an-integrated-approach-to-analyzing-the-behavior-and-function-of-the-glycocalyx-using-both-electron-and-fluorescence-intravital-microscopy
#16
Hanae Kataoka, Akira Ushiyama, Yoshihiro Akimoto, Sachie Matsubara, Hayato Kawakami, Takehiko Iijima
BACKGROUND: The endothelial surface layer (ESL) regulates vascular permeability to maintain fluid homeostasis. The glycocalyx (GCX), which has a complex and fragile ultrastructure, is an important component of the ESL. Abnormalities of the GCX have been hypothesized to trigger pathological hyperpermeability. Here, we report an integrated in vivo analysis of the morphological and functional properties of the GCX in a vital organ. METHODS: We examined the behavior of the ESL and GCX, using both electron microscopy (EM) and intravital microscopy (IVM)...
May 11, 2017: Anesthesia and Analgesia
https://www.readbyqxmd.com/read/28504320/photosensitization-priming-of-tumor-microenvironments-improves-delivery-of-nanotherapeutics-via-neutrophil-infiltration
#17
Dafeng Chu, Xinyue Dong, Qi Zhao, Jingkai Gu, Zhenjia Wang
Remodeling of tumor microenvironments enables enhanced delivery of nanoparticles (NPs). This study shows that direct priming of a tumor tissue using photosensitization rapidly activates neutrophil infiltration that mediates delivery of nanotherapeutics into the tumor. A drug delivery platform is comprised of NPs coated with anti-CD11b antibodies (Abs) that target activated neutrophils. Intravital microscopy demonstrates that the movement of anti-CD11b Abs-decorated NPs (NPs-CD11b) into the tumor is mediated by neutrophil infiltration induced by photosensitization (PS) because the systemic depletion of neutrophils completely abolishes the nanoparticle tumor deposition...
May 15, 2017: Advanced Materials
https://www.readbyqxmd.com/read/28503576/sirtuin-2-regulates-microvascular-inflammation-during-sepsis
#18
Nancy Buechler, Xianfeng Wang, Barbara K Yoza, Charles E McCall, Vidula Vachharajani
Objective. Sepsis and septic shock, the leading causes of death in noncoronary intensive care units, kill more than 200,000/year in the US alone. Circulating cell-endothelial cell interactions are the rate determining factor in sepsis inflammation. Sirtuin, a seven-member family of proteins (SIRT1-7), epigenetically controls inflammation. We have studied the roles of SIRTs 1, 3, and 6 in sepsis previously. In this project, we studied the role of SIRT2 on sepsis-related inflammation. Methods. Sepsis was induced in C57Bl/6 (WT), SIRT2 knockout (SIRT2KO), and SIRT2 overexpressing (SIRT2KI) mice by cecal ligation and puncture (CLP)...
2017: Journal of Immunology Research
https://www.readbyqxmd.com/read/28490813/leptin-receptor-antagonism-of-inkt-cell-function-a-novel-strategy-to-combat-multiple-myeloma
#19
M Favreau, E Menu, D Gaublomme, K Vanderkerken, S Faict, K Maes, E De Bruyne, S Govindarajan, M Drennan, S Van Calenbergh, X Leleu, L Zabeau, J Tavernier, K Venken, D Elewaut
A hallmark of bone marrow changes with aging is the increase in adipocyte composition, but how this impacts development of multiple myeloma (MM) is unknown. Here, we report the role of the adipokine leptin as master regulator of anti-myeloma tumor immunity by modulating the invariant natural killer T (iNKT) cell function. A marked increase in serum leptin levels and leptin receptor (LR) expression on iNKT cells in MM patients and the 5T33 murine MM model was observed. MM cells and leptin synergistically counteracted anti-tumor functionality of both murine and human iNKT cells...
May 11, 2017: Leukemia: Official Journal of the Leukemia Society of America, Leukemia Research Fund, U.K
https://www.readbyqxmd.com/read/28473106/from-good-to-bad-intravital-imaging-of-the-hijack-of-physiological-processes-by-cancer-cells
#20
REVIEW
Saskia J E Suijkerbuijk, Jacco van Rheenen
Homeostasis of tissues is tightly regulated at the cellular, tissue and organismal level. Interestingly, tumor cells have found ways to hijack many of these physiological processes at all the different levels. Here we review how intravital microscopy techniques have provided new insights into our understanding of tissue homeostasis and cancer progression. In addition, we highlight the different strategies that tumor cells have adopted to use these physiological processes for their own benefit. We describe how visualization of these dynamic processes in living mice has broadened to our view on cancer initiation and progression...
May 1, 2017: Developmental Biology
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