scholarly journals Intravital Fluorescence Microscopy: A Novel Tool for the Study of the Interaction ofStaphylococcus aureuswith the Microvascular Endothelium In Vivo

2005 ◽  
Vol 191 (3) ◽  
pp. 435-443 ◽  
Author(s):  
Matthias W. Laschke ◽  
Sylvain Kerdudou ◽  
Mathias Herrmann ◽  
Michael D. Menger
Keyword(s):  
Fluorescence Microscopy ◽  
Intravital Fluorescence Microscopy ◽  
Microvascular Endothelium

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

4D in in vivo 2-photon laser scanning fluorescence microscopy with sample motion in 6 degrees of freedom

2011 ◽  
Vol 200 (1) ◽  
pp. 47-53 ◽  
Author(s):  
Sabine Scheibe ◽  
Mario M. Dorostkar ◽  
Christian Seebacher ◽  
Rainer Uhl ◽  
Frank Lison ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Laser Scanning ◽  
Degrees Of Freedom ◽  
Sample Motion ◽  
6 Degrees Of Freedom

scholarly journals In vivo spectral and fluorescence microscopy comparison of microvascular function after treatment with OXi4503, Sunitinib and their combination in Caki-2 tumors

2014 ◽  
Vol 5 (6) ◽  
pp. 1965 ◽  
Author(s):  
Jennifer A. Lee ◽  
Nikolett M. Biel ◽  
Raymond T. Kozikowski ◽  
Dietmar W. Siemann ◽  
Brian S. Sorg
Keyword(s):  
Fluorescence Microscopy ◽  
Microvascular Function

Polymeric/Inorganic Multifunctional Nanoparticles for Simultaneous Drug Delivery and Visualization

2010 ◽  
Vol 1257 ◽  
Author(s):  
Andrea Fornara ◽  
Alberto Recalenda ◽  
Jian Qin ◽  
Abhilash Sugunan ◽  
Fei Ye ◽  
...  
Keyword(s):  
Drug Delivery ◽  
Fluorescence Microscopy ◽  
Contrast Agents ◽  
Gold Nanorods ◽  
In Vitro Cytotoxicity ◽  
In Vivo Studies ◽  
Multifunctional Nanoparticles ◽  
Cytotoxicity Studies

AbstractNanoparticles consisting of different biocompatible materials are attracting a lot of interest in the biomedical area as useful tools for drug delivery, photo-therapy and contrast enhancement agents in MRI, fluorescence and confocal microscopy. This work mainly focuses on the synthesis of polymeric/inorganic multifunctional nanoparticles (PIMN) based on biocompatible di-block copolymer poly(L,L-lactide-co-ethylene glycol) (PLLA-PEG) via an emulsion-evaporation method. Besides containing a hydrophobic drug (Indomethacin), these polymeric nanoparticles incorporate different visualization agents such as superparamagnetic iron oxide nanoparticles (SPION) and fluorescent Quantum Dots (QDs) that are used as contrast agents for Magnetic Resonance Imaging (MRI) and fluorescence microscopy together. Gold Nanorods are also incorporated in such nanostructures to allow simultaneous visualization and photodynamic therapy. MRI studies are performed with different loading of SPION into PIMN, showing an enhancement in T2 contrast superior to commercial contrast agents. Core-shell QDs absorption and emission spectra are recorded before and after their loading into PIMN. With these polymeric/inorganic multifunctional nanoparticles, both MRI visualization and confocal fluorescence microscopy studies can be performed. Gold nanorods are also synthesized and incorporated into PIMN without changing their longitudinal absorption peak usable for lased excitation and phototherapy. In-vitro cytotoxicity studies have also been performed to confirm the low cytotoxicity of PIMN for further in-vivo studies.

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