Name: retrograde loading of nerves, tracts, and spinal roots with fluorescent dyes
Uploaded: 2012-04-19T13:00:00
Description: Watch this Scientific Journal Video about Retrograde Loading of Nerves, Tracts, and Spinal Roots with Fluorescent Dyes at JoVE.com

<p class="jove_content"&gt; Dit werk werd ondersteund door de intramurale programma van de National Institutes of Neurologische Aandoeningen en Stroke in de National Institutes of Health. Wij zouden ook graag Dr George Mentis bedanken voor zijn eerdere bijdragen aan de methode en de gegevens in figuur 3.</p

Fluorescerende dextranen zijn gebruikt als anatomische gereedschappen en voor het afbeelden van neuronale activiteit 1-4. Velden et al.., (2009) 4 publiceerde een protocol voor het toepassen van ionen-en voltage-sensitieve kleurstoffen om axonale stukken met een focus op het ruggenmerg als modelsysteem. Hier beschrijven we een meer gedetailleerde procedure voor de toepassing van TL-en / of ion-gevoelige kleurstof om de snede ventrale wortels, dorsale wortels of een neuronale kanaal van het...

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<p class="jove_content"&gt; We beschrijven hier een eenvoudige en kosteneffectieve protocol voor de invoering van kleurstoffen in de neuronen, de zenuwen en spinale traktaten. Deze methode omvat het blootstellen geïdentificeerd anatomische trajecten een zeer geconcentreerde kleurstof oplossing voor de duur van het laden. Dit resulteert in een retrograde etikettering van de target site met weinig achtergrond in vergelijking met micro-injecties, bad toepassing en elektroporatie technieken. Echter, de werkwijze beperkt tot sites in het zenuwstels...

<table border="1"&gt;<tbody&gt;<tr&gt;<td colspan="2"<strong&gt; Materiaalnaam</strong</td&gt;<td<strong&gt; Bedrijf</strong</td&gt;<td<strong&gt; Catalogus nummer</strong</td&gt;<td<strong&gt; Commentaar</strong</td&gt;</tr&gt;<tr&gt;<td rowspan="2"&gt; Slangen</td&gt;<td&gt; PE90 (IDxOD: 0,034 &quot;x0.050&quot;; muur Thikness: 0.008 &quot;)</td&gt;<td&gt; Clay Adams Brand Intermedic</td&gt;<td&gt; 427.421</td&gt;<td&gt; Voor type I en type II pipetten</td&gt;</tr&gt;<tr&gt;<td&gt; NSF-51 (IDxOD: 1/16x1/8; muur Thikness: 1/32)</td&gt;<td&gt; PharMed BPT, Cole-Parmer</td&gt;<td&gt; AY242002</td&gt;<td&gt; Voor Flexibele buis</td&gt;</tr&gt;<tr&gt;<td&gt; Spuit</td&gt;<td&gt; 1 ml insuline spuit U-100</td&gt;<td&gt; Becton Dickenson</td&gt;<td&gt; 329.650</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Naald</td&gt;<td&gt; 19G x1-1/2 &quot;Metal Hub Naald</td&gt;<td&gt; MONOJECT</td&gt;<td&gt; 200.136</td&gt;<td&gt; Aansluiten van Type II pipet op de spuit</td&gt;</tr&gt;<tr&gt;<td&gt;</td&gt;<td&gt; Alcohol Lamp</td&gt;<td&gt;</td&gt;<td&gt;</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Afsluitkraan</td&gt;<td&gt; Drie-wegkraan met mannelijke Luer Slip-adapter</td&gt;<td&gt; Baxter Healthcare Corp</td&gt;<td&gt; 2C6241</td&gt;<td&gt; Gebruik met spuit</td&gt;</tr&gt;<tr&gt;<td rowspan="3"&gt; Houder</td&gt;<td&gt; H-1 elektrodehouder</td&gt;<td&gt; Narishige</td&gt;<td&gt; H-1/12</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Magnet stand</td&gt;<td&gt; Narishige</td&gt;<td&gt; GJ-8</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Micromanipulator</td&gt;<td&gt; Narishige</td&gt;<td&gt; M-3333</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Pincet</td&gt;<td&gt; W &amp; T-DUMONT Zwitserse 00632-11</td&gt;<td&gt; DUMONT</td&gt;<td&gt; JF-5 TC</td&gt;<td&gt;</td&gt;</tr&gt;</tbody&gt;</table&gt;<p class="jove_content"&gt; * Dat zijn voorgestelde materialen. Kan worden vervangen door elke compatibele producten</p

retrograde loading of nerves, tracts, and spinal roots with fluorescent dyes

Retrograde etikettering van neuronen is een standaard anatomische methode 1,2 die ook is gebruikt om calcium-en voltage-sensitieve kleurstoffen te laden in neuronen 3-6. Over het algemeen worden de kleurstoffen toegepast als vaste kristallen, of door lokale druk injectie met glazen pipetten. Dit kan echter leiden tot een verdunning van de kleurstof en een verminderde intensiteit van de etikettering, in het bijzonder wanneer meerdere uren nodig zijn voor kleurstofdiffusie. Hier tonen een eenvoudige en goedkope methode voor het inbrengen fluorescentie en ion-gevoelige kleurstoffen in neuronen met een polyethyleen zuig pipet gevuld met de kleurstofoplossing. Deze methode biedt een betrouwbare wijze voor handhaving van een hoge concentratie van de kleurstof in contact met axons gedurende het laden.

Watch this Scientific Journal Video about Retrograde Loading of Nerves, Tracts, and Spinal Roots with Fluorescent Dyes at JoVE.com

Retrograde Loading of Nerves, Tracts, and Spinal Roots with Fluorescent Dyes

Wij beschrijven een eenvoudige en goedkope methode voor het inbrengen hoge concentratie van fluorescerende en calcium-gevoelige kleurstoffen in neuronen of neuronale kanaal met een polyethyleen zuig pipet.

Retrograde Laden van zenuwen, Tracts, en Spinal Roots met fluorescente kleurstoffen

Retrograde labeling of neurons is a standard anatomical method1,2 that has also been used to load calcium and voltage-sensitive dyes into neurons3-6. ...

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