CD4 + Tリンパ球キャプチャHIVのための使い捨てマイクロ流体チップを用いた

Protocol

抗CD4 + biotinlayted表面とマイクロ流体チャネルを準備します。 シリンジポンプとマイクロ流体チャネルに5ml /分でサンプルの血液10mlを注入する。 PBSで20分間20ml /分で血を洗い流す。 15分間5ul/minで希釈FACS溶解溶液を（PBSで原液から10％に希釈）を注入。 15分間インキュベートする。 20分間5ml /分で一次CD3 +抗体溶液を（PBSで原液から10％に希釈）を注入。 …

Materials

Material Name	Type	Company	Catalogue Number	Comment
PDMS microchannel				Fabricated by SU-8 mold
Blood				Donated blood from normal human subject. Mass General Hospital, 10 X high density WBC or Brigham and Women’s Hospital.
PBS	Buffer	Invitrogen	14190	1x Dulbecco’s Phosphated Buffered Saline.
FACS lysing solution		Becton Dickinson	349202	Diluted to 10% from stock solution with PBS.
Anti-CD4+ biotinlayted Ab		Beckman Coulter	IM0704	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.
goat anti-hCD3delta	pAB	Santa Cruz Biotechnology	sc-1128	Diluted to 10% from stock solution with PBS
rabbit anti-goat IgG PE		Santa Cruz Biotechnology	sc-3755	Secondary Ab fro anti-CD3 Ab, diluted to 10% from stock solution with PBS
anti-human CD4+		Becton Dickinson	557695	Alexa Fluor 488 labeled, diluted to 20% from stock solution with PBS.
DAPI		Invitrogen	D1306	4’,6-dianidino-2-phenylindole, dihydrochloride solution, diluted to 0.2% from stock solution with PBS.