<ol><li> Mikroakışkan kanal anti-CD4 <sup>+</sup> biotinlayted yüzey hazırlayın. </li><li> 5 ml / dk 10 ml numune kan şırınga pompası ile mikroakışkan kanal içine enjekte edilir. </li><li> PBS ile 20 dakika için 20 ml / dak kan yıkayın. </li><li> 15 dakika için 5ul/min seyreltilmiş FACS parçalama solüsyonu (PBS ile stok solüsyonu% 10 sulandırılmış) enjekte edilir. </li><li> 15 dakika boyunca inkübe. </li><li> 5 ml / dk 20 dk birincil CD3 <sup>+</sup> antikor solüsyonu (PBS ile stok solüsyonu% 10 sulandırılmış) enjekte edilir. </li><li> 2 saat süreyle inkübe edin. </li><li> 5 ml / dk 15 dk PBS ile kanal yıkayın. </li><li> 5 ml / dk, aynı zamanda 20 dakika boyunca% 20 AF488 CD4 <sup>+</sup> antikor,% 0.2 DAPI stok solüsyonu,% 10 ikincil CD3 <sup>+</sup> antikor enjekte edilir. </li><li> 20 dakika boyunca inkübe. </li><li> 5 ml / dk 15 dk kanalı PBS ile yıkayın. </li><li> Floresan mikroskop altında hücrelerin sayısı: <ol><li> Beyaz (optik) - Siyah ve beyaz: hücre ve toz </li><li> Yalnızca UV (DAPI) - Mavi: hücre </li><li> Blue (CD4 <sup>+)</sup> - Yeşil: T-lenfosit ve monosit </li><li> Sadece Yeşil (CD3 <sup>+)</sup> - Kırmızı: T-lenfosit </li></ol></li></ol>

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		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>PDMS microchannel</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Fabricated by SU-8 mold</td>
</tr>
<tr>
<td>Blood</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Donated blood from normal human subject. Mass General Hospital, 10 X high density WBC or Brigham and Women&#x2019;s Hospital.</td>
</tr>
<tr>
<td>PBS</td>
<td>Buffer</td>
<td>Invitrogen</td>
<td>14190</td>
<td>1x Dulbecco&#x2019;s Phosphated Buffered Saline.</td>
</tr>
<tr>
<td>FACS lysing solution</td>
<td>&nbsp;</td>
<td>Becton Dickinson</td>
<td>349202</td>
<td>Diluted to 10% from stock solution with PBS.</td>
</tr>
<tr>
<td>Anti-CD4+ biotinlayted Ab</td>
<td>&nbsp;</td>
<td>Beckman Coulter</td>
<td>IM0704</td>
<td>Surface prepared with Anti-CD4+ biotinlayted Ab, diluted to 10% from stock solution with PBS/BSA after mixing with 0.2mg stock and 1ml deionized water. </td>
</tr>
<tr>
<td>goat anti-hCD3delta</td>
<td>pAB</td>
<td>Santa Cruz Biotechnology</td>
<td>sc-1128</td>
<td>Diluted to 10% from stock solution with PBS</td>
</tr>
<tr>
<td>rabbit anti-goat IgG PE</td>
<td>&nbsp;</td>
<td>Santa Cruz Biotechnology</td>
<td>sc-3755</td>
<td>Secondary Ab fro anti-CD3 Ab, diluted to 10% from stock solution with PBS</td>
</tr>
<tr>
<td>anti-human CD4+</td>
<td>&nbsp;</td>
<td>Becton Dickinson</td>
<td>557695</td>
<td>Alexa Fluor 488 labeled, diluted to 20% from stock solution with PBS. </td>
</tr>
<tr>
<td>DAPI</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>D1306</td>
<td>4&#x2019;,6-dianidino-2-phenylindole, dihydrochloride solution, diluted to 0.2% from stock solution with PBS.</td>
</tr>		</tbody>
		</table>
		</div>
	

cd4+ t-lymphocyte capture using a disposable microfluidic chip for hiv

Watch this Scientific Journal Video about CD4+ T-Lymphocyte Capture Using a Disposable Microfluidic Chip for HIV at JoVE.com

CD4 + T-Lenfosit Yakalama HIV için Disposable mikroakışkan Chip