<ol><li> Prepare canais microfluídicos com anti-CD4 <sup>+</sup> biotinlayted superfície. </li><li> Injectar 10 ml de amostra de sangue de 5 ml / min no canal microfluídicos com bomba de seringa. </li><li> Lavar o sangue de 20 ml / min para 20 min com PBS. </li><li> Injectar a solução diluída de lise FACS (diluído a 10% da solução estoque com PBS) em 5ul/min por 15 min. </li><li> Incubar por 15 min. </li><li> Injetar CD3 <sup>+</sup> solução de anticorpos primários (diluído a 10% da solução estoque com PBS) a 5 ml / min para 20 min. </li><li> Incubar por 2 horas. </li><li> Lave canal com PBS, 5 ml / min para 15 min. </li><li> Injectar 20% AF488 CD4 <sup>+</sup> de anticorpos, solução estoque 0,2% DAPI, e 10% CD3 <sup>+</sup> anticorpo secundário a 5 ml / min para 20 min, ao mesmo tempo. </li><li> Incubar por 20 min. </li><li> Lave o canal com PBS, 5 ml / min para 15 min. </li><li> Contagem de células ao microscópio fluorescente: <ol><li> White (óptica) - Preto e branco: célula e poeira </li><li> UV (DAPI) - Blue: única célula </li><li> Azul (CD4 <sup>+)</sup> - Verde: linfócitos T e monócitos </li><li> Verde (CD3 <sup>+)</sup> - Vermelho: linfócitos T só </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-linfócitos Captura Usando um chip descartável microfluídicos para HIV