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Intravital microscopy

Jochen-Frederick Hernekamp, Florian Neubrech, Tomke Cordts, Volker J Schmidt, Ulrich Kneser, Thomas Kremer
PURPOSE: The clinical course after major burns is characterized by microcirculatory changes and consecutive capillary leakage. However, current clinical monitoring does not properly assess microcirculation, whereas macrohemodynamic changes are continuously evaluated. Here, we assess if macrohemodynamic and microhemodynamic parameters after burn trauma are correlated in a rat model. METHODS: Burn plasma harvested from donor rats 4 hours after thermal injury (30% total body surface area, 100 °C water, 12 seconds) was administered intravenously to healthy animals during 2 hours of intravital microscopy (burn group [BG])...
October 17, 2016: Annals of Plastic Surgery
Edwin D Hawkins, Delfim Duarte, Olufolake Akinduro, Reema A Khorshed, Diana Passaro, Malgorzata Nowicka, Lenny Straszkowski, Mark K Scott, Steve Rothery, Nicola Ruivo, Katie Foster, Michaela Waibel, Ricky W Johnstone, Simon J Harrison, David A Westerman, Hang Quach, John Gribben, Mark D Robinson, Louise E Purton, Dominique Bonnet, Cristina Lo Celso
It is widely accepted that complex interactions between cancer cells and their surrounding microenvironment contribute to disease development, chemo-resistance and disease relapse. In light of this observed interdependency, novel therapeutic interventions that target specific cancer stroma cell lineages and their interactions are being sought. Here we studied a mouse model of human T-cell acute lymphoblastic leukaemia (T-ALL) and used intravital microscopy to monitor the progression of disease within the bone marrow at both the tissue-wide and single-cell level over time, from bone marrow seeding to development/selection of chemo-resistance...
October 17, 2016: Nature
P Larsson, I Alwis, B Niego, M Sashindranath, P Fogelstrand, M C L Wu, L Glise, M Magnusson, M Daglas, N Bergh, S P Jackson, R L Medcalf, S Jern
BACKGROUND: The endogenous fibrinolytic system has rarely been considered as a target to prevent thrombotic disease. Tissue plasminogen activator (t-PA) is potently increased by histone deacetylase (HDAC) inhibitors in endothelial cells in vitro, but whether this translates into increased vascular t-PA production and an enhanced fibrinolytic capacity in vivo is unknown. OBJECTIVES: The aim of this study was to determine if the HDAC inhibitor valproic acid (VPA) stimulates production of t-PA in the vasculature of mice, and further, if VPA pre-treatment affects fibrin deposition and clot formation after mechanical vessel injury...
October 5, 2016: Journal of Thrombosis and Haemostasis: JTH
Sylvia M Bardet, Lynn Carr, Malak Soueid, Delia Arnaud-Cormos, Philippe Leveque, Rodney P O'Connor
Despite the biomedical advances of the last century, many cancers including glioblastoma are still resistant to existing therapies leaving patients with poor prognoses. Nanosecond pulsed electric fields (nsPEF) are a promising technology for the treatment of cancer that have thus far been evaluated in vitro and in superficial malignancies. In this paper, we develop a tumor organoid model of glioblastoma and apply intravital multiphoton microscopy to assess their response to nsPEFs. We demonstrate for the first time that a single 10 ns, high voltage electric pulse (35-45 kV/cm), collapses the perfusion of neovasculature, and also alters the diameter of capillaries and larger vessels in normal tissue...
October 4, 2016: Scientific Reports
Almudena Ortega Gomez, Melanie Salvermoser, Jan Rossaint, Robert Pick, Janine Brauner, Patricia Lemnitzer, Jessica Tilgner, Renske de Jong, Remco T A Megens, Janina Jamasbi, Yvonne Döring, Christine T Pham, Christoph Scheiermann, Wolfgang Siess, Maik Drechsler, Christian Weber, Jochen Grommes, Alexander Zarbock, Barbara Walzog, Oliver Soehnlein
BACKGROUND: -Therapeutic targeting of arterial leukocyte recruitment in the context of atherosclerosis has been disappointing in clinical studies. Reasons for such failures include the lack of knowledge of arterial-specific recruitment patterns. Here we establish the importance of the Cathepsin G (CatG) in the context of arterial myeloid cell recruitment. METHODS: -Intravital microscopy of the carotid artery, the jugular vein and cremasteric arterioles and venules in Apoe(-/-) and CatG-deficient mice Apoe(-/-)Ctsg(-/-)) mice was employed to study site-specific myeloid cell behavior after high fat diet (HFD) feeding or TNF stimulation...
September 22, 2016: Circulation
Sung Hoon Baik, Seokjo Kang, Sung Min Son, Inhee Mook-Jung
Pathological hallmarks of Alzheimer's disease (AD) include extracellularly accumulated amyloid β (Aβ) plaques and intracellular neurofibrillary tangles in the brain. Activated microglia, brain-resident macrophages, are also found surrounding Aβ plaques. The study of the brain of AD mouse models revealed that Aβ plaque formation is completed by the consolidation of newly generated plaque clusters in vicinity of existed plaques. However, the dynamics of Aβ plaque formation, growth and the mechanisms by which microglia contribute to Aβ plaque formation are unknown...
September 23, 2016: Glia
M W Laschke, M D Menger
The dorsal skinfold chamber is a rodent model for non-invasive microcirculatory analyses of striated muscle and skin tissue throughout an observation period of 2-3 weeks. In combination with intravital fluorescence microscopy, this model allows the quantitative assessment of dynamic processes such as inflammation, angiogenesis, vascular remodelling and microcirculation. Accordingly, the dorsal skinfold chamber is increasingly used for preclinical research in tissue engineering and regenerative medicine. This includes studies on biocompatibility, vascularisation and incorporation of medical implants and artificial tissue constructs...
2016: European Cells & Materials
Muhamad Marlini, Ayako Mabuchi, Beth L Mallard, Ngatiman Hairulhisyam, Sachiko Akashi-Takamura, Jacquie L Harper, Antony M Wheatley
Liver regeneration is delayed in mice with a defective TLR4 (C3H/HeJ) but is normal in TLR4 knockouts (TLR4(-/-) ). Here, we investigated the possible involvement of structural and hemodynamic changes in vivo in the underlying mechanism. In LPS-sensitive (C3H/HeN, C57BL/6) and LPS-insensitive (C3H/HeJ, TLR4(-/-) ) mice, a 70% partial hepatectomy (PH) was performed under inhalational anesthesia. At days 3 and 7 after PH, the hepatic microcirculation was interrogated using intravital microscopy. Delayed liver regeneration was confirmed in C3H/HeJ, but not in C3H/HeN, C57BL/6 (WT) or TLR4(-/-) mice by liver weight-to-body-weight ratio, percentage of PCNA-positive cells and mitotic index data...
September 16, 2016: Experimental Physiology
Galit H Frydman, Anna Le, Felix Ellett, Julianne Jorgensen, James G Fox, Ronald G Tompkins, Daniel Irimia
Neutrophils are traditionally regarded as the "first responders" of the immune system. However, recent observations revealed that platelets often respond earlier to recruit and activate neutrophils within sites of injury and inflammation. Currently, platelet-neutrophil interactions are studied by intravital microscopy. Although such studies provide exceptional, physiologic in vivo data, they are also laborious and have low throughput. To accelerate platelet-neutrophil interaction studies, we have developed and optimized an ex vivo microfluidic platform with which the interactions between platelets and moving neutrophils are measured at single-cell level in precise conditions and with high throughput...
September 14, 2016: Journal of Leukocyte Biology
Gabriel Cerqueira Alves Costa, Adriana Coelho Soares, Marcos Horácio Pereira, Nelder Figueiredo Gontijo, Maurício Roberto Viana Sant'Anna, Ricardo Nascimento Araujo
Ornithodoros rostratus is an argasid tick and its importance is based on its hematophagy and the resulting transmission of pathogens such as Rickettsia rickettsii and Coxiella burnetii unto its vertebrate hosts. In the face of a lack of physiological studies related to hematophagy in argasid ticks, this paper aims to identify and characterize the events that occur throughout the feeding by O. rostratus on live hosts. Electrical signals and alterations on the feeding site were monitored using intravital microscopy and electromyography...
September 13, 2016: Journal of Experimental Biology
Thanushiyan Poobalasingam, David Salman, Henry Li, Joana Alçada Costa, Charlotte H Dean
Our understanding of lung biology can be greatly enhanced by studying embryonic and postnatal lung development, and the perturbations which occur during disease. Imaging techniques provide a unique insight into these processes. A wide variety of imaging techniques have been used to study the lungs at various stages of development and disease, ranging from histological stains to more novel techniques such as single plane illumination microscopy (SPIM), intravital microscopy (IVM), and micro-computed tomography (micro-CT)...
September 14, 2016: Histology and Histopathology
Marc Lepeltier, Florence Appaix, Yuan Yuan Liao, Frédéric Dumur, Jérôme Marrot, Tangui Le Bahers, Chantal Andraud, Cyrille Monnereau
A tris-cyclometalated iridium complex that bears two ligands functionalized by peripheral carbazole groups combines an intense solid state emission and a significant two-photon absorption cross section in the near-infrared. After incorporation into a physiological micellar suspension, it can be used for the intravital two-photon fluorescence microscopy of cerebral vasculature.
October 3, 2016: Inorganic Chemistry
Kyoohyun Kim, Kibaek Choe, Inwon Park, Pilhan Kim, YongKeun Park
Intravital microscopy is an essential tool that reveals behaviours of live cells under conditions close to natural physiological states. So far, although various approaches for imaging cells in vivo have been proposed, most require the use of labelling and also provide only qualitative imaging information. Holographic imaging approach based on measuring the refractive index distributions of cells, however, circumvent these problems and offer quantitative and label-free imaging capability. Here, we demonstrate in vivo two- and three-dimensional holographic imaging of circulating blood cells in intact microcapillaries of live mice...
2016: Scientific Reports
Mykhailo Vladymyrov, Jun Abe, Federica Moalli, Jens V Stein, Akitaka Ariga
The development of multi-photon intravital microscopy, in particular two-photon microscopy (2PM), has been a breakthrough technique for deep-tissue imaging of dynamic cell behavior inside live organisms and has substantially advanced the field of immunology. However, intravital time-lapse imaging over prolonged time periods is complicated by slow tissue drifts caused by vital activity, leading to shifting fields of views and making the acquired image sequence partially or completely unanalyzable. To solve this issue, we have established a system that performs continuous drift offset correction in real time using fine pattern matching during 2PM acquisition...
November 2016: Journal of Immunological Methods
Daisuke Nakano, Akira Nishiyama
The number of people being diagnosed with end-stage renal disease is increasing globally. Therapeutic options to slow or halt the progression of kidney disease are limited and are not always successful, despite the increasing body of research and number of basic scientific reports in this field. Further studies are required to investigate new approaches to renal pathophysiology. State of the art optical imaging is a powerful tool used to non-invasively observe the pathophysiology of small animals and has the potential to elucidate the unknown mechanisms of renal disease and aid in our understanding of the disease...
September 2016: Journal of Pharmacological Sciences
Victor Naumenko, Craig Jenne, Douglas J Mahoney
The development of intravital microscopy has provided unprecedented capacity to study the tumor microenvironment in live mice. The dynamic behavior of cancer, stromal, vascular, and immune cells can be monitored in real time, in situ, in both primary tumors and metastatic lesions, allowing treatment responses to be observed at single cell resolution and therapies tracked in vivo. These features provide a unique opportunity to elucidate the cellular mechanisms underlying the biology and treatment of cancer. We describe here a method for imaging the microenvironment of subcutaneous tumors grown in mice using intravital microscopy...
2016: Methods in Molecular Biology
Liane Babes, Paul Kubes
Intravital microscopy has evolved into an invaluable technique to study the complexity of tumors by visualizing individual cells in live organisms. Here, we describe a method for employing intravital spinning disk confocal microscopy to picture high-resolution tumor-stroma interactions in real time. We depict in detail the surgical procedures to image various tumor microenvironments and different cellular components in the liver.
2016: Methods in Molecular Biology
Lian Willetts, David Bond, Konstantin Stoletov, John D Lewis
Metastasis, or the spread of cancer cells from a primary tumor to distant sites, is the leading cause of cancer-associated death. Metastasis is a complex multi-step process comprised of invasion, intravasation, survival in circulation, extravasation, and formation of metastatic colonies. Currently, in vitro assays are limited in their ability to investigate these intricate processes and do not faithfully reflect metastasis as it occurs in vivo. Traditional in vivo models of metastasis are limited by their ability to visualize the seemingly sporadic behavior of where and when cancer cells spread (Reymond et al...
2016: Methods in Molecular Biology
Florian S Frueh, Thomas Später, Nicole Lindenblatt, Maurizio Calcagni, Pietro Giovanoli, Claudia Scheuer, Michael D Menger, Matthias W Laschke
Full-thickness skin defects can be covered with dermal skin substitutes in combination with split-thickness skin grafts. However, slow vascularization of the matrices bears the risk of wound infection and extends the length of hospitalization. To overcome these problems, we describe a promising vascularization strategy. Green fluorescent protein(+) adipose tissue-derived microvascular fragments (ad-MVF) were isolated from epididymal fat pads of C57BL/6-Tg(CAG-EGFP)1Osb/J mice. ad-MVF were seeded on collagen-glycosaminoglycan matrices, which were implanted into full-thickness skin defects in the dorsal skinfold chamber of wild-type C57BL/6 mice...
August 26, 2016: Journal of Investigative Dermatology
Anna-Maria Szczesniak, Richard F Porter, James T Toguri, Joanna Borowska-Fielding, Simon Gebremeskel, Anuja Siwakoti, Brent Johnston, Christian Lehmann, Melanie E M Kelly
Proliferative vitreoretinopathy (PVR) can develop after ocular trauma or inflammation and is a common complication of surgery to correct retinal detachment. Currently, there are no pharmacological treatments for PVR. Cannabinoids acting at cannabinoid 2 receptor (CB2R) can decrease inflammation and fibrosis. The objective of this study was to examine the anti-inflammatory actions of CB2R as a candidate novel therapeutic target in experimental PVR. PVR was induced by intravitreal injection of dispase in wild-type (WT) and CB2R genetic knockout (CB2R(-/-)) mice...
August 25, 2016: Neuropharmacology
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