pH controlled staining of CD4+ and CD19+ cells within functionalized microfluidic channel |
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| Authors: | Mariangela Mortato Laura Blasi Giovanna Barbarella Simona Argentiere Giuseppe Gigli |
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| Institution: | 1.Superior School ISUFI, University of Salento, via Arnesano, I-73100 Lecce, Italy;2.NNL CNR-Institute of Nanoscience, via Arnesano, I-73100 Lecce, Italy;3.CNR - ISOF, I-40129 Bologna, Italy;4.Fondazione Filarete Srl, Viale Ortles 22/4, 20139 Milano, Italy;5.Department of Innovation Engineering, University of Salento, via Arnesano, I-73100 Lecce, Italy;6.Istituto Italiano di Tecnologia, IIT-Lecce, Via Barsanti, 73010 Lecce, Italy |
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| Abstract: | Herein proposed is a simple system to realize hands-free labeling and simultaneous detection of two human cell lines within a microfluidic device. This system was realized by novel covalent immobilization of pH-responsive poly(methacrylic acid) microgels onto the inner glass surface of an assembled polydimethylsiloxane/glass microfluidic channel. Afterwards, selected thiophene labeled monoclonal antibodies, specific for recognition of CD4 antigens on T helper/inducer cells and CD19 antigens on B lymphocytes cell lines, were encapsulated in their active state by the immobilized microgels. When the lymphocytes suspension, containing the two target subpopulations, was flowed through the microchannel, the physiological pH of the cellular suspension induced the release of the labeled antibodies from the microgels and thus the selective cellular staining. The selective pH-triggered staining of the CD4- and CD19-positive cells was investigated in this preliminary experimental study by laser scanning confocal microscopy. This approach represents an interesting and versatile tool to realize cellular staining in a defined module of lab-on-a-chip devices for subsequent detection and counting. |
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