Direct observation of liposome uptake by leukocytes in vivo in skin blood vessels using intravital fluorescence microscopy

2000 ◽  
Author(s):  
Jean-Marie Devoisselle ◽  
Serge R. Mordon ◽  
Sylvie Begu ◽  
Thomas Desmettre
Keyword(s):  
Fluorescence Microscopy ◽  
Blood Vessels ◽  
Direct Observation ◽  
Intravital Fluorescence Microscopy ◽  
Skin Blood Vessels

In vivobehaviour of long-circulating liposomes in blood vessels in hamster inflammation and septic shock models-use of intravital fluorescence microscopy

Luminescence ◽  
2001 ◽  
Vol 16 (2) ◽  
pp. 73-78 ◽  
Author(s):  
Jean-Marie Devoisselle ◽  
Sylvie Begu ◽  
Corine Tourné-Péteilh ◽  
Thomas Desmettre ◽  
Serge Mordon
Keyword(s):  
Septic Shock ◽  
Fluorescence Microscopy ◽  
Blood Vessels ◽  
Intravital Fluorescence Microscopy ◽  
Shock Models

scholarly journals Injection of Evans blue dye to fluorescently label and image intact vasculature

BioTechniques ◽  
2020 ◽  
Author(s):  
Samuel E Honeycutt ◽  
Lori L O'Brien
Keyword(s):  
Fluorescence Microscopy ◽  
Blood Vessels ◽  
Evans Blue ◽  
Blue Dye ◽  
Vascular Networks ◽  
Fluorescent Properties ◽  
Evans Blue Dye ◽  
Health And Disease ◽  
Disease Understanding

Blood vessels perform critical functions in both health and disease. Understanding how vessels form, pattern and respond to damage is essential. However, labeling and imaging the vasculature to ascertain these properties can be difficult and time-consuming. Here, the authors present a novel methodology for rapidly and efficiently labeling whole vascular networks in vivo by exploiting the fluorescent properties of Evans blue. By combining the labeling with fluorescence microscopy, this method enables visualization of whole tissue vasculature for a fraction of the time and cost compared with traditional methods.

Abstract 28: Serial in Vivo Imaging of Thrombus Inflammation Predicts Venous Thrombus Resolution

2012 ◽  
Vol 32 (suppl_1) ◽  
Author(s):  
Chase W Kessinger ◽  
Jason R McCarthy ◽  
Charles P Lin ◽  
Peter K Henke ◽  
Farouc A Jaffer
Keyword(s):  
Fluorescence Microscopy ◽  
In Vivo Imaging ◽  
Macrophage Infiltration ◽  
Intravital Fluorescence Microscopy ◽  
Targeted Nanoparticles ◽  
Macrophage Activity ◽  
Vein Thrombosis ◽  
Deep Vein ◽  
In Vivo Assessment

Introduction: Inflammation following deep vein thrombosis (DVT) critically modulates thrombus resolution. However it is unknown whether the degree of inflammation in vivo predicts the magnitude of thrombus resolution. Here, we utilize serial multichannel intravital fluorescence microscopy (IVFM) to spatially map and quantify macrophage infiltration in thrombus, and then to determine the degree of thrombus resolution in vivo . Methods: C57Bl/6 mice (n=5) underwent topical ferric chloride injury to induce femoral venous thrombosis. On day 3, mice were i.v. injected with macrophage-targeted nanoparticles (CLIO-AF555, ex/em 555/565nm). On day 4, survival IVFM of thrombus macrophages was performed. In addition, venography was concomitantly performed (pre-injection of FITC-dextran 45 minutes before IVFM, MW 2000 kD, 490/520nm) to quantify DVT area and length in vivo at day 4 and 6. IVFM image analysis was performed using ImageJ. Mid-luminal z-stacks (40 μm thickness) were analyzed. Thrombus ROIs were outlined using FITC-dextran generated angiograms, and used calculating macrophage target-to-background ratios (TBR) and thrombus area and length. Contralateral sham femoral veins were imaged as controls. Imaging was followed by histopathology and fluorescence microscopy of the cryosectioned tissue. Results: At day 4, the baseline femoral DVT area and length were 0.22±0.08mm 2 and 1.29±0.16mm (mean±sd). The day 4 macrophage activity was noted to be heterogeneous with peripheral infiltration of the thrombus, and occasionally with some deeper penetration into the thrombus interior. At day 6, the thrombus area and length were reduced compared to day 4 (p<0.005 for both). The day 4 thrombus macrophage TBR significantly predicted the degree of thrombus reduction (correlation of initial macrophage TBR and Δthrombus area, r=0.97, p=0.001). Conclusion: This serial in vivo assessment of murine DVT resolution demonstrates that degree of macrophage infiltration at day 4 predicts the magnitude of thrombus resolution at day 6. Molecular imaging of DVT inflammation in vivo could provide new insights into DVT resolution and the development of the post-thrombotic syndrome, and also in evaluating inflammatory-modulating therapies for improved DVT resolution.

Hepatic microcirculatory failure after ischemia and reperfusion: improvement with ATP-MgCl2 treatment

1985 ◽  
Vol 248 (6) ◽  
pp. H804-H811 ◽  
Author(s):  
M. G. Clemens ◽  
P. F. McDonagh ◽  
I. H. Chaudry ◽  
A. E. Baue
Keyword(s):  
Fluorescence Microscopy ◽  
Unit Area ◽  
Hepatic Function ◽  
Ischemia And Reperfusion ◽  
Intravital Fluorescence Microscopy ◽  
Hepatic Microcirculation ◽  
Hepatic Ischemia ◽  
Beneficial Effects ◽  
Valuable Method

Hepatic ischemia followed by reflow results in a myriad of metabolic and circulatory derangements that may eventually result in liver failure and death. In the present experiments we have used the technique of intravital fluorescence microscopy with fluoroscein isothiocyanate conjugated to bovine serum albumin as the intravascular fluorochrome to study the effects of ischemia and reperfusion on the hepatic microcirculation in vivo. Total hepatic ischemia was produced for 90 min to the left and median lobes of pentobarbital-anesthetized rats. After ischemia, reflow was allowed for 2 h. Three groups were studied: sham-ischemia controls and rats treated with either 1 ml saline or 12.5 mumol ATP-MgCl2 in 1-ml volume at the beginning of reflow. Although control rats exhibited stable microcirculation throughout the experiment, in saline-treated rats the number of perfused centrilobular areas and perfused sinusoids per unit area on the surface of the liver was decreased to approximately 50 and 40% of sham-ischemia controls, respectively. However, in rats treated with ATP-MgCl2 the density of perfused centrilobular areas and perfused sinusoids was 86 and 80% of sham-ischemia controls, respectively. From these results we conclude that intravital fluorescence microscopy is a potentially valuable method for the study of the hepatic microcirculation in vivo. Moreover, the results with ATP-MgCl2 treatment indicate that its effect on the microcirculation is a major factor in its beneficial effects on hepatic function after ischemia and reflow.

In vivo analysis of angiogenesis in endometriosis-like lesions by intravital fluorescence microscopy

2005 ◽  
Vol 84 ◽  
pp. 1199-1209 ◽  
Author(s):  
Matthias W. Laschke ◽  
Antje Elitzsch ◽  
Brigitte Vollmar ◽  
Michael D. Menger
Keyword(s):  
Fluorescence Microscopy ◽  
Intravital Fluorescence Microscopy ◽  
In Vivo Analysis

In Vivo Spin Trapping of Glyceryl Trinitrate–Derived Nitric Oxide in Rabbit Blood Vessels and Organs

Circulation ◽  
1995 ◽  
Vol 92 (7) ◽  
pp. 1876-1882 ◽  
Author(s):  
Alexander Mülsch ◽  
Peter Mordvintcev ◽  
Eberhard Bassenge ◽  
Frank Jung ◽  
Bernd Clement ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Glyceryl Trinitrate ◽  
Blood Vessels ◽  
Spin Trapping ◽  
Rabbit Blood

A High-Affinity Human Antibody That Targets Tumoral Blood Vessels

Blood ◽  
1999 ◽  
Vol 94 (1) ◽  
pp. 192-198 ◽  
Author(s):  
Lorenzo Tarli ◽  
Enrica Balza ◽  
Francesca Viti ◽  
Laura Borsi ◽  
Patrizia Castellani ◽  
...  
Keyword(s):  
Blood Vessels ◽  
Small Cell Lung Carcinoma ◽  
Antibody Fragment ◽  
Human Colon ◽  
Experimental Models ◽  
Human Antibody ◽  
Cell Lung Carcinoma ◽  
High Affinity ◽  
Biodistribution Studies

Angiogenesis is a characteristic feature of many aggressive tumors and of other relevant disorders. Molecules capable of specifically binding to new-forming blood vessels, but not to mature vessels, could be used as selective vehicles and would, therefore, open diagnostic and therapeutic opportunities. We have studied the distribution of the ED-B oncofetal domain of fibronectin, a marker of angiogenesis, in four different tumor animal models: the F9 murine teratocarcinoma, SKMEL-28 human melanoma, N592 human small cell lung carcinoma, and C51 human colon carcinoma. In all of these experimental models we observed accumulation of the fibronectin isoform containing the ED-B domain around neovascular structures when the tumors were in the exponentially growing phase, but not in the slow-growing phase. Then we performed biodistribution studies in mice bearing a subcutaneously implanted F9 murine teratocarcinoma, using a high-affinity human antibody fragment (L19) directed against the ED-B domain of fibronectin. Radiolabeled L19, but not an irrelevant anti-lysozyme antibody fragment (D1.3), efficiently localizes in the tumoral vessels. The maximal dose of L19 accumulated in the tumor was observed 3 hours after injection (8.2% injected dose per gram). By virtue of the rapid clearance of the antibody fragment from the circulation, tumor-to-blood ratios of 1.9, 3.7, and 11.8 were obtained at 3, 5, and 24 hours, respectively. The tumor-targeting performance of L19 was not dose-dependent in the 0.7 to 10 μg range of injected antibody. The integral of the radioactivity localized in tumoral vessels over 24 hours was greater than 70-fold higher than the integral of the radioactivity in blood over the same time period, normalized per gram of tissue or fluid. These findings quantitatively show that new-forming blood vessels can selectively be targeted in vivo using specific antibodies, and suggest that L19 may be of clinical utility for the immunoscintigraphic detection of angiogenesis in patients.

scholarly journals An in vivo model allowing continuous observation of human vascular formation in the same animal over time

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yohei Tsukada ◽  
Fumitaka Muramatsu ◽  
Yumiko Hayashi ◽  
Chiaki Inagaki ◽  
Hang Su ◽  
...  
Keyword(s):  
Blood Vessel ◽  
Blood Vessels ◽  
Human Blood ◽  
Molecular Mechanisms ◽  
Human Tumor ◽  
Continuous Observation ◽  
In Vivo Models ◽  
Cranial Window ◽  
Pathological Conditions

AbstractAngiogenesis contributes to numerous pathological conditions. Understanding the molecular mechanisms of angiogenesis will offer new therapeutic opportunities. Several experimental in vivo models that better represent the pathological conditions have been generated for this purpose in mice, but it is difficult to translate results from mouse to human blood vessels. To understand human vascular biology and translate findings into human research, we need human blood vessel models to replicate human vascular physiology. Here, we show that human tumor tissue transplantation into a cranial window enables engraftment of human blood vessels in mice. An in vivo imaging technique using two-photon microscopy allows continuous observation of human blood vessels until at least 49 days after tumor transplantation. These human blood vessels make connections with mouse blood vessels as shown by the finding that lectin injected into the mouse tail vein reaches the human blood vessels. Finally, this model revealed that formation and/or maintenance of human blood vessels depends on VEGFR2 signaling. This approach represents a useful tool to study molecular mechanisms of human blood vessel formation and to test effects of drugs that target human blood vessels in vivo to show proof of concept in a preclinical model.

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