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

Dette arbejde blev støttet af en bevilling af &quot;Deutsche Forschungsgemeinschaft&quot; til SU Eisenhardt (EI 866/1-1).

1. Anæstesi og overvågning <ol><li> Passende nationale og institutionelle etik skal være på plads, før du udfører dyreforsøg. Efter godkendelse fra den etiske komité bedøver Sprague Dawley rotter med en kropsvægt fra 120 til 180 g. Lever 2 til 3 vol% isofluran til en plexiglas kasse via isofluran fordamper og placere rotten inde. </li><li> Så snart det passende niveau af anæstesi er opnået (manglende reaktion på tåen eller hale knibe) rotte vægtes og barberedes på den ventrale cervikale område. </li>...

<ol>
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Leukocyt-endothelial interaktion, produktion af reaktive ilt arter og aktivering af komplementsystemet er de vigtigste funktioner i IRI-induceret væv dysfunktion. 26 mikrocirkulationen af det angrebne væv betragtes som integralet stedet for den inflammatoriske debut. Bortset fra ex vivo eksperimenter såsom flow kammer analyser 27, 28 er det obligatorisk at give veletablerede modeller for intravital billedbehandling til yderligere at evaluere in vivo relevans. Selvom IRI er blev...

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

Iskæmi-reperfusionsskade (IRI) har været impliceret i en lang række patologiske tilstande, såsom cerebralt slagtilfælde, myocardieinfarkt, intestinal iskæmi såvel som efter transplantationen og kardiovaskulær kirurgi. En Reperfusion af tidligere iskæmisk væv, mens afgørende for forebyggelse af irreversibel vævsskade, fremkalder overdreven inflammation af det påvirkede væv. Tilgrænsende til produktion af reaktive oxygenarter, aktivering af komplementsystemet og forøget mikrovaskulær permeabilitet, aktivering af leukocytter er en af de vigtigste aktører i patologiske kaskade af inflammatorisk vævsbeskadigelse under reperfusion. 2, 3 leukocytaktivering er en flertrinsproces, der består rullende, faste adhæsion og transmigration og er medieret af et komplekst samspil mellem adhæsionsmolekyler som reaktion på kemoattraktanter såsom komplementfaktorer, kemokiner eller blodplade-aktiverende faktor. 4 &lt;p class = &quot;jove_content&quot;&gt; Mens leukocyt rulning i postkapillare venuler hovedsageligt medieres ved interaktion af selectiner 5 med deres modparter ligander, faste adhæsion af leukocytter til endotelet, er selectin-styret via binding til intercellulære adhæsionsmolekyler (ICAM) og vaskulær cellulære adhæsionsmolekyler (VCAM). 6, 7 Guldstandarden til in vivo observation af leukocyt-endotel-interaktion er en teknik intravital mikroskopi, først beskrevet i 1968. 8 Selvom forskellige modeller af IRI (iskæmi-reperfusionsbeskadigelse), er blevet beskrevet til forskellige organer, 9-12 kun få er egnede til direkte visualisering af leukocytrekruttering i mikrovaskulære leje på et højt niveau af billedkvaliteten. 8 Vi her fremme den digitale intravital epifluorescens-mikroskopi af postkapillære venulen i cremasteric mikrocirkulationaf rotte 13 som en bekvem metode til kvalitativt og kvantitativt at analysere leukocytrekruttering for IRI-forskning i tværstribet muskelvæv og giver en detaljeret manual for udførelse af teknikken. Vi yderligere at belyse almindelige faldgruber og give nyttige tips, der skal gøre det muligt for læseren at virkelig værdsætte, og sikkert udføre metoden. I en trinvis protokol, som vi skildrer, hvordan du kommer i gang med åndedræt kontrolleret anæstesi under tilstrækkelig overvågning til at holde dyret fast bedøvet i længere perioder. Vi så beskriver cremasteric præparatet som en tynd flad plade for fremragende optisk opløsning og giver en protokol for leukocyt billeddannelse i IRI, der er blevet veletableret i vores 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 epifluorescens-mikroskopi af postkapillære venuler i den cremasteric mikrocirkulation er en bekvem metode til at få indblik i leukocyt-endotel-interaktion<em&gt; In vivo</em&gt; I iskæmi-reperfusionsskade (IRI) af tværstribet muskelvæv. Vi her give en detaljeret protokol til sikkert at udføre teknikken og diskutere dens anvendelsesmuligheder og begrænsninger.

Realtid Digital Imaging af leukocyt-endotel Interaktion med iskæmi-reperfusionsskade (IRI) af rotte Cremaster Muscle

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

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Medicine

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 Tværgående Rectus abdominis Myocutaneous Flap: En Rat Model of Myocutaneous iskæmisk reperfusionsbeskadigelse

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Iskæmireperfusion Model akut nyreskade og Post Injury Fibrose i mus

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 ...

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

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Induktion Iskæmi-reperfusionsskade i mus Øre Skin for Intravital multifotonexcitation Imaging af immunrespons

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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Evaluering af koronar flow reserve efter myokardieiskæmi reperfusion hos rotter

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

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En præklinisk rotte model for studiet af iskæmi-reperfusion skade i rekonstruktiv mikrokirurgi

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

Forsinket Intramyokardie levering af stamceller efter Iskæmi Reperfusion Skade i en Murine Model

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 Iskæmi-Reperfusion Skade ved at simulere hæmodynamiske ændringer i rottelevertransplantation Model

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