Name: real-time digital imaging of leukocyte-endothelial interaction in ischemia-reperfusion injury (iri) of the rat cremaster muscle
Uploaded: 2012-08-05T14:00:00
Description: Watch this Scientific Journal Video about Real-time Digital Imaging of Leukocyte-endothelial Interaction in Ischemia-reperfusion Injury IRI of the Rat Cremaster Muscle at JoVE.com

Dit werk werd ondersteund door een subsidie ​​van de &quot;Deutsche Forschungsgemeinschaft&quot; om SU Eisenhardt (EI 866/1-1).

1. Anesthesie en Monitoring <ol><li> Passende nationale en institutionele ethiek moet zijn voordat het uitvoeren van dierproeven. Na goedkeuring van de ethische commissie Anesthetize mannelijke Sprague Dawley ratten met een lichaamsgewicht van 120 tot 180 g. Lever 2 tot 3 vol% isofluraan een plexiglazen box via isofluraan vaporizer en plaats de rat naar binnen. </li><li> Zodra het juiste niveau van de anesthesie wordt bereikt (gebrek aan reactie op teen of staart knijpen) de rat wordt gewogen en geschoren aan de ventr...

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Leukocyten-endotheliale interactie, de productie van reactieve zuurstof soorten en activering van het complementsysteem zijn de belangrijkste kenmerken van de IRI-geïnduceerde weefsel disfunctie. 26 De microcirculatie van het aangetaste weefsel wordt beschouwd als de integrale site voor de ontstekingsreactie ontstaan. Naast de ex vivo experimenten zoals stroomkamer assays 27, 28 is het verplicht om gevestigde modellen van intravitale beeldvorming te verstrekken aan verdere beoordeling van...

<table border="1"><tbody><tr><td> Naam van de apparatuur: </td><td> Bedrijf: </td><td> Artikelnummer: </td><td> Opmerkingen: </td></tr><tr><td> Forene 100% (V / V) </td><td> Abt </td><td> B506 </td><td> API isofluraan </td></tr><tr><td> Textiel binnenwerk Suture </td><td> Serag Weissner </td><td> OC108000 </td><td></td></tr><tr><td> Portex Fine Boring Polythene Tubing </td><td> Smiths Medical </td><td> 800/100/100 </td><td> 0,28 mm binnendiameter </td></tr><tr><td> 0,9% zoutoplossing </td><td> Fresinus Kabi </td><td> 808771 </td><td></td></tr><tr><td> Change-A-tip luxe cauterisatie kit </td><td> Bovie Medical </td><td> DEL1 </td><td></td></tr><tr><td> Abbocath-T-14G </td><td> Venisystems </td><td> G713- A01 </td><td> als optische buis </td></tr><tr><td> Servo Ventilator 900C </td><td> Maquet </td><td></td><td> als dier ventialtor </td></tr><tr><td> Logische drukomvormer </td><td> Smiths Medical </td><td> MX1960 </td><td></td></tr><tr><td> Sirecust 404 Monitor </td><td> Siemens </td><td></td><td></td></tr><tr><td> ABL 700 Benchtop Analyzer </td><td> Radiometer </td><td></td><td> voor bloed gasmeting </td></tr><tr><td> Verwarming pad </td><td> Effenberger </td><td> 8319 </td><td></td></tr><tr><td> Aluminium fase </td><td> Alfun </td><td> AW7022 </td><td></td></tr><tr><td> Chirurgische microscoop OPMI 6-SDFC </td><td> Carl Zeiss </td><td></td><td></td></tr><tr><td> Microchirurgische instrumenten lab set </td><td> S &amp; T </td><td> 767 </td><td></td></tr><tr><td> Biemer schip clip </td><td> Diener </td><td> 64.562 </td><td></td></tr><tr><td> Het toepassen tang </td><td> Diener </td><td> 64.568 </td><td> voor Biemer schip clip </td></tr><tr><td> Rhodamine 6G </td><td> Sigma-Aldrich </td><td> R4127 </td><td></td></tr><tr><td> Vaseline witte DAB </td><td> Winthrop </td><td> 2726853 </td><td></td></tr><tr><td> Cover bril </td><td></td><td></td><td> 32x32 mm </td></tr><tr><td></td><td></td><td></td><td> Intravitale setup </td></tr><tr><td> Zeiss Axio Scope A-1 MAT </td><td> Carl Zeiss </td><td> 490036 </td><td> epifluorescentiemicroscoop </td></tr><tr><td> 470 nm LED </td><td> Carl Zeiss </td><td> 423052 </td><td> fluorescentie lichtbron </td></tr><tr><td> Colibri 2-systeem </td><td> Carl Zeiss </td><td> 423052 </td><td></td></tr><tr><td> W Plan-Apochromat 20x / 1,0 DIC </td><td> Carl Zeiss </td><td> 421452 </td><td> onderdompeling in water doel </td></tr><tr><td> Axiocam MRM Rev 3 FireWire </td><td> Carl Zeiss </td><td> 426509 </td><td> hoge resolutie digitale camera </td></tr><tr><td> Axio visie LE-software </td><td> Carl Zeiss </td><td> 410130 </td><td> te gebruiken voor offline analyse </td></tr></tbody></table>

real-time digital imaging of leukocyte-endothelial interaction in ischemia-reperfusion injury (iri) of the rat cremaster muscle

Ischemie-reperfusie schade (IRI) is betrokken bij een groot scala aan pathologische aandoeningen zoals cerebrale beroerte, hartinfarct, darmischemie en na de transplantatie en cardiovasculaire chirurgie. Een Reperfusie van eerder ischemisch weefsel, terwijl het van essentieel belang voor het voorkomen van onomkeerbare weefselbeschadiging, ontlokt veel ontsteking van het aangetaste weefsel. Naast de productie van zuurstofradicalen, activering van het complement systeem verhoogd microvasculaire permeabiliteit, de activering van leukocyten is een van de voornaamste actoren de pathologische cascade van inflammatoire weefselbeschadiging reperfusie. 2, 3 leukocytactivatiesyndroom een complex proces bestaat rollend, stevige hechting zielsverhuizing en wordt bemiddeld door een complex interactie tussen adhesiemoleculen reactie op chemoattractants zoals complementfactoren, chemokines, en bloedplaatjes-activerende factor 4. &lt;p class = &quot;jove_content&quot;&gt; Terwijl rollen van leukocyten in postcapillaire venules voornamelijk gemedieerd wordt door de interactie van selectines 5 met hun liganden tegen, stevige hechting van leukocyten aan het endotheel is selectine gecontroleerde via binding aan intercellular adhesion molecules (ICAM) en vasculaire cellulaire adhesiemoleculen (VCAM). 6, 7 Gouden standaard voor de in vivo waarnemingen van leukocyt-endotheliale interactie is de techniek van intravitaal microscopie eerst beschreven in 1968 8. Hoewel de verschillende modellen van IRI (ischemie-reperfusie schade) zijn beschreven voor verschillende organen, 9-12 slechts enkele zijn geschikt voor directe visualisatie van leukocyt rekrutering in de microvasculaire bed op een hoog niveau van beeldkwaliteit. 8 Wij hier bevorderen digitale intravitaal epifluorescente microscopie van de postcapillaire venule de cremasteric microcirculatievan de rat 13 als een handige methode om leukocyten werving kwalitatief en kwantitatief te analyseren voor de IRI-onderzoek in dwarsgestreept spierweefsel en een gedetailleerde handleiding voor het vervullen van de techniek. We illustreren verder voorkomende valkuilen en geven nuttige tips, die in staat moet stellen de lezer echt te waarderen, en veilig uit te voeren van de methode. In een stap voor stap protocol waarin we beschrijven hoe te beginnen met de ademhaling gecontroleerd anesthesie in voldoende controle om het dier stevig onder narcose te houden voor een langere periode van tijd. Daarna beschrijven we de cremasteric preparaat als een dunne vlakke plaat voor een uitstekende optische resolutie en een protocol voor leukocyten beeldvorming in IRI, dat goed is vastgelegd in onze laboratoria.

Watch this Scientific Journal Video about Real-time Digital Imaging of Leukocyte-endothelial Interaction in Ischemia-reperfusion Injury IRI of the Rat Cremaster Muscle at JoVE.com

Real-time Digital Imaging of Leukocyte-endothelial Interaction in Ischemia-reperfusion Injury IRI of the Rat Cremaster Muscle

Digitale intravitale epifluorescentie microscopie van postcapillaire venulen in de cremasteric microcirculatie is een handige methode om inzicht te krijgen in leukocyten-endotheliale interactie<em&gt; In vivo</em&gt; In ischemie-reperfusie schade (IRI) van dwarsgestreept spierweefsel. Wij hier een gedetailleerd protocol om veilig uit te voeren de techniek en de toepassingen en beperkingen te bespreken.

Real-time Digital Imaging van leukocyt-endotheliale Interactie in ischemie-reperfusie schade (IRI) van de Rat cremasterspier

Ischemia-reperfusion injury (IRI) has been implicated in a large array of pathological conditions such as cerebral stroke, myocardial infarction, ...

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Real-time Digital Imaging of Leukocyte-endothelial Interaction in Ischemia-reperfusion Injury (IRI) of the Rat Cremaster Muscle

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Ischemia-reperfusion injury (IRI) occurs when there is transient hypoxia due to the obstruction of blood flow (ischemia) followed by a subsequent ...

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Een pre-klinische rat model voor de studie van ischemie-reperfusie letsel in reconstructieve microchirurgie

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many ...

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