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

Detta arbete stöddes av ett bidrag av &quot;Deutsche Forschungsgemeinschaft&quot; för att SU Eisenhardt (EI 866/1-1).

1. Anestesi och övervakning <ol><li> Lämpliga nationella och institutionella etik bör vara på plats innan du utför djurförsök. Efter godkännande från etikkommittén bedöva Sprague Dawley råttor med en kroppsvikt från 120 till 180 gram. Leverera 2 till 3 vol% isofluran till Plexiglas boxen via isofluran förångare och placera råttan insidan. </li><li> Så snart en lämplig nivå av anestesi uppnås (brist på reaktion på tå eller svans nypa) är råttan viktas och rakade på ventrala cervikala området....

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Leukocyt-endoteliala interaktion, produktion av reaktiva syrespecies och aktivering av komplementsystemet är de viktigaste egenskaperna hos IRI-inducerad vävnad dysfunktion. 26 mikrocirkulationen av den påverkade vävnaden betraktas som integralen platsen för den inflammatoriska debut. Bortsett från ex vivo experiment som analyser flödeskammare 27, 28 är det obligatoriskt att tillhandahålla väl etablerade modeller av intravital avbildning för att ytterligare utvärdera in viv...

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

Ischemi-reperfusionsskada (IRI) har implicerats i ett stort arrangemang av patologiska tillstånd, såsom cerebral stroke, myokardinfarkt, intestinal ischemi såväl som efter transplantationskirurgi och kardiovaskulär kirurgi. 1 Reperfusion av tidigare ischemisk vävnad, medan väsentliga för förhindrande av irreversibel vävnadsskada, framkallar överdrivet inflammation i angripna vävnaden. Intill produktion av reaktiva syrespecies, aktivering av komplementsystemet och ökad mikrovaskulär permeabilitet, är aktiveringen av leukocyter en av de principiella aktörer i patologiska kaskad av inflammatorisk vävnadsskada under reperfusion. 2, 3 är leukocytaktivering en flerstegsprocess som består valsningsriktningen, starka adhesion och och medieras av en komplex interaktion mellan adhesionsmolekyler som svar på kemoattraktanter såsom komplementfaktorer, kemokiner, eller blodplättsaktiverande faktor. 4 &lt;p klass = &quot;jove_content&quot;&gt; Medan leukocytrullning i postkapillära venoler huvudsakligen medieras genom interaktion av selektiner 5 med deras motsvarigheter ligander, fast adhesion av leukocyter till endotelet är selektin-styrd via bindning till intercellulära adhesionsmolekyler (ICAM) och vaskulär cellulära adhesionsmolekyler (VCAM) 6, 7 Gyllene standarden för in vivo-observation av leukocyt-endoteliala interaktion är tekniken för intravital mikroskopi, som först beskrevs 1968. 8 Fastän olika modeller av IRI (ischemi-reperfusionsskada) har beskrivits för olika organ, 9-12 endast ett fåtal är lämpliga för direkt visualisering av leukocytrekrytering i den mikrovaskulära bädden på en hög nivå av bildkvalitet. 8 Vi främjar här digitala intravital epifluorescensmikroskopi av postkapillära venolen i cremasteric mikrocirkulationenav rått 13 som en bekväm metod för att kvalitativt och kvantitativt analysera leukocytrekrytering för IRI-forskning i tvärstrimmig muskelvävnad och ger en detaljerad handbok för att åstadkomma tekniken. Vi illustrerar ytterligare gemensamma fallgropar och ger användbara tips som ska hjälpa läsaren att verkligen uppskatta, och säkert utföra metoden. I ett steg för steg protokoll vi skildra hur du kommer igång med andningen kontrollerad anestesi under tillräcklig övervakning för att hålla djuret ordentligt sövd under längre perioder. Vi beskriver sedan cremasteric beredningen som en tunn platt ark för enastående optisk upplösning och ger ett protokoll för leukocyt avbildning i IRI som har väl etablerade i våra laboratorier.

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

Digital intravital epifluorescensmikroskopi av postkapillära venoler i cremasteric mikrocirkulationen är en bekväm metod för att få insikter i leukocyt-endoteliala interaktionen<em&gt; In vivo</em&gt; I ischemi-reperfusionsskada (IRI) i tvärstrimmig muskelvävnad. Vi erbjuder här ett detaljerat protokoll för att säkert utföra tekniken och diskutera dess tillämpningar och begränsningar.

Real-time Digital bildbehandling av leukocyt-endotel interaktion i ischemi-reperfusionsskada (IRI) i råtta kremastermuskeln

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

In situ Transverse Rectus Abdominis Myocutaneous Flap: A Rat Model of Myocutaneous Ischemia Reperfusion Injury

In situ Transverse Rectus Abdominis Myocutaneous Flap: A Rat Model of Myocutaneous Ischemia Reperfusion Injury

In situ Tvärgående Rectus abdominis Myocutaneous Flap: En Rat modell av Myocutaneous ischemireperfusionsskada

Free tissue transfer is the gold standard of reconstructive surgery to repair complex defects not amenable to local options or those requiring composite ...

Ischemia-reperfusion Model of Acute Kidney Injury and Post Injury Fibrosis in Mice

Ischemi-reperfusion modell av akut Kidneyskadan och efter skada Fibros i möss

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often ...

A Murine Model of Myocardial Ischemia-reperfusion Injury through Ligation of the Left Anterior Descending Artery

En mus modell av ischemi-reperfusion skada genom Ligation i vänstra främre nedåt Artery

Acute or chronic myocardial infarction (MI) are cardiovascular events resulting in high morbidity and mortality. Establishing the pathological mechanisms ...

Induction and Assessment of Ischemia-reperfusion Injury in Langendorff-perfused Rat Hearts

Induktion och Bedömning av ischemi-reperfusion skada i Langendorff-perfusion Rat Hearts

The biochemical events surrounding ischemia reperfusion injury in the acute setting are of great importance to furthering novel treatment options for ...

Murine Model of Intestinal Ischemia-reperfusion Injury

Murina Modell av Intestinal ischemi-reperfusionsskada

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, ...

Inducing Ischemia-reperfusion Injury in the Mouse Ear Skin for Intravital Multiphoton Imaging of Immune Responses

Induktion ischemi-reperfusionsskada i musen Ear Skin för Intravital multifoton avbildning av immunsvar

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

Evaluation of Coronary Flow Reserve After Myocardial Ischemia Reperfusion in Rats

Utvärdering av Koronarflödesreserv efter myokardischemi Refusion hos råttor

Coronary artery disease is the leading cause of death worldwide. After an acute myocardial infarction, early and successful myocardial intervention via ...

A Pre-clinical Rat Model for the Study of Ischemia-reperfusion Injury in Reconstructive Microsurgery

En preklinisk råtta modell för studiet av ischemia-reperfusion skada i rekonstruktiv microsurgery

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

Delayed Intramyocardial Delivery of Stem Cells after Ischemia Reperfusion Injury in a Murine Model

Fördröjd Intramyokardiell leverans av stamceller efter Ischemia Reperfusion Skada i en Murine Modell

There is significant interest in the use of stem cells (SCs) for the recovery of cardiac function in individuals with myocardial injuries. Most commonly, ...

Organ Ischemia-Reperfusion Injury by Simulating Hemodynamic Changes in Rat Liver Transplant Model

Organ ischemi-reperfusion skada genom att simulera hemodynamic förändringar i råtta lever transplantation modell

Orthotopic liver transplantation (OLT) in rats is a tried and proven animal model used for preoperative, intraoperative, and postoperative studies, ...