Name: characterization of the isolated, ventilated, and instrumented mouse lung perfused with pulsatile flow
Uploaded: 2011-04-29T17:00:00
Description: Watch this Scientific Journal Video about Characterization of the Isolated, Ventilated, and Instrumented Mouse Lung Perfused with Pulsatile Flow at JoVE.com

Dit onderzoek werd ondersteund door de National Institutes of Health subsidie ​​R01HL086939 (NCC).

In dit protocol tonen we een geïsoleerde, geventileerde, doorbloed muis long preparaat dat eerder is gebruikt om de impact van de longembolie en chronische hypoxie op de pulserende pulmonale vasculaire druk-flow relaties (Tuchscherer, Webster, en Chesler, 2006 kwantificeren; Tuchscherer et al. ., 2007). In het kort worden de muis longen operatief geïsoleerd van het omliggende weefsel, geplaatst in een verwarmde kamer (IL-1; Harvard Apparatuur, Holliston, MA) en geventileerde (Ventilatoire Control Module (VCM)-R met ti...

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Kritische stappen in de operatie Het is van cruciaal belang dat de zorg wordt genomen bij het snijden van de ribbenkast uit de buurt van de longen. De longen moeten volledig worden vrijgemaakt en onbevangen door het omliggende weefsel tijdens de inflatie, maar niet beschadigd tijdens het proces van isolatie. Het gebruik van een plat voorwerp, zoals de achterkant van de tang kan worden gebruikt om weg te houden van de longen van de borstwand, zodat er een duidelijk pad voor de schaar te knippen....

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		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>1 ml syringe</td>
<td>&nbsp;</td>
<td>Fisher Sci</td>
<td>14-829-10F</td>
<td>&nbsp;</td>
<tr>
<td>10 ml syringe</td>
<td>&nbsp;</td>
<td>Fisher Sci</td>
<td>14-823-2A </td>
<td>&nbsp;</td>
<tr>
<td>60 ml syringe</td>
<td>&nbsp;</td>
<td>Fisher Sci</td>
<td>13-689-8 </td>
<td>&nbsp;</td>
<tr>
<td>RPMI with GLN 6/PK</td>
<td>&nbsp;</td>
<td>Fisher Sci</td>
<td>MT10040CV </td>
<td>&nbsp;</td>
<tr>
<td>Bottle Top Filters</td>
<td>&nbsp;</td>
<td>Fisher Sci</td>
<td>09-761-57</td>
<td>&nbsp;</td>
<tr>
<td>Ficoll PM 70</td>
<td>&nbsp;</td>
<td>Sigma-Aldrich</td>
<td>F2878-100g</td>
<td>&nbsp;</td>
<tr>
<td>Heparin</td>
<td>&nbsp;</td>
<td>Sigma-Aldrich</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>Y27632</td>
<td>&nbsp;</td>
<td>Sigma-Aldrich</td>
<td>Y0503</td>
<td>&nbsp;</td>
<tr>
<td>Angled Ball Iris scissors</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>14109-09</td>
<td>&nbsp;</td>
<tr>
<td>Vannas Spring Scissors - 4mm Blades</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>15018-10</td>
<td>&nbsp;</td>
<tr>
<td>Fine Iris Scissors - straight</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>14106-09</td>
<td>&nbsp;</td>
<tr>
<td>Dumont #5/45 Forceps</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>11251-35</td>
<td>&nbsp;</td>
<tr>
<td>Dumont Medical Biology Forceps</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>11254-20</td>
<td>&nbsp;</td>
<tr>
<td>Lauda E100 ECO-line 003 </td>
<td>&nbsp;</td>
<td>VWR</td>
<td>Comparable to Lauda-Brinkmann E-103, 62400-922</td>
<td>&nbsp;</td>
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<td>IL-1 Isolated perfused mouse lung system </td>
<td>&nbsp;</td>
<td>Harvard Apparatus </td>
<td>739904 </td>
<td>&nbsp;</td>
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<td>Blood Pressure Transducer P75 for PLUGSYS Module</td>
<td>&nbsp;</td>
<td>Harvard Apparatus</td>
<td>730020</td>
<td>&nbsp;</td>
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<td>TS410 Flow Modules</td>
<td>&nbsp;</td>
<td>Transonic </td>
<td>TS410</td>
<td>&nbsp;</td>
<tr>
<td>ME 4 PXN Precision PXN Inline Flowsensors</td>
<td>&nbsp;</td>
<td>Transonic</td>
<td>ME 4 PXN</td>
<td>&nbsp;</td>
<tr>
<td>Cole-Parmer Multi-Syringe Pumps</td>
<td>&nbsp;</td>
<td>Cole-Parmer </td>
<td>EW-74900-20</td>
<td>&nbsp;</td>
<tr>
<td>Nembutal 50MG/ML 20ML Vial</td>
<td>&nbsp;</td>
<td>Amatheon</td>
<td>&nbsp;</td>
<td>&nbsp;</td>		</tbody>
		</table>
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characterization of the isolated, ventilated, and instrumented mouse lung perfused with pulsatile flow

De geïsoleerde, geventileerde en geïnstrumenteerde muis longen voorbereiding maakt stabiel en pulserende pulmonale vasculaire druk-flow relaties te meten met onafhankelijke controle over de pulmonale arteriële doorstroming, het debiet golfvorm, luchtwegdruk en linker atrium druk. Pulmonale vasculaire weerstand wordt berekend op basis van multi-point, constante druk-flow curves; pulmonale vasculaire weerstand wordt berekend op basis van pulsatiele druk-flow curves verkregen bij een bereik van frequenties. Zoals nu klinisch erkend, impedantie is een superieure maat voor rechter ventrikel afterload dan de weerstand omdat het ook de effecten van vasculaire compliance, die niet verwaarloosbaar zijn, vooral in de pulmonale circulatie. Drie belangrijke statistieken van de impedantie - de nul hertz impedantie Z 0, de karakteristieke impedantie Z C, en de index van golf reflectie R W - inzicht geven in distale arteriële dwarsdoorsnede beschikbaar voor flow, proximale arteriële stijfheid en de upstream-downstream impedantie mismatch, respectievelijk. Alle resultaten die in geïsoleerde, geventileerde en doorbloed longen zijn onafhankelijk van het sympathische zenuwstelsel toon, volume de status en de effecten van de anesthesie. We hebben gebruik gemaakt van deze techniek om de impact van de longembolie en chronische hypoxie op de weerstand en impedantie te kwantificeren, en om onderscheid te maken tussen sites van de actie (dat wil zeggen, proximale versus distale) van vasoactieve stoffen en ziekten met behulp van de druk afhankelijkheid van Z-C. Bovendien, wanneer deze technieken worden gebruikt met de longen van genetisch gemanipuleerde stammen van muizen, kunnen de effecten van moleculair niveau defecten op pulmonale vasculaire structuur en functie worden bepaald.

Watch this Scientific Journal Video about Characterization of the Isolated, Ventilated, and Instrumented Mouse Lung Perfused with Pulsatile Flow at JoVE.com

Characterization of the Isolated, Ventilated, and Instrumented Mouse Lung Perfused with Pulsatile Flow

Het volgende protocol schetst het proces van isoleren, ventileren en instrumenteren muis longen stabiel of pulsatiele pulmonale vasculaire druk-flow relaties maatregel om de effecten van de bloedstroom, luchtstroom, luchtweg veranderingen en vasculaire veranderingen op rechter ventrikel afterload kwantificeren.

Karakterisering van de geïsoleerde, geventileerde en Instrumented Mouse Lung geperfuseerd met pulsatiele flow

The isolated, ventilated and instrumented mouse lung preparation allows steady and pulsatile pulmonary vascular pressure-flow relationships to be measured ...

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