An inkjet-printed polysaccharide matrix for on-chip sample preparation in point-of-care cell counting chambers |
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Authors: | Xichen Zhang Dorothee Wasserberg Christian Breukers Bridgette J. Connell Pauline J. Schipper Joost van Dalum Ellen Baeten Dorine van den Blink Andries C. Bloem Monique Nijhuis Annemarie M. J. Wensing Leon W. M. M. Terstappen Markus Beck |
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Affiliation: | Medical Cell Biophysics, MIRA Institute for Biomedical Technology and Technical Medicine, Faculty of Science and Technology, University of Twente, PO Box 217, Enschede 7500 AE The Netherlands.; University Medical Center Utrecht, Department of Medical Microbiology, Virology, Heidelberglaan 100, 3584 CX Utrecht The Netherlands ; University Medical Center Utrecht, Laboratory of Translational Immunology, Section Diagnostics, Heidelberglaan 100, 3584 CX Utrecht The Netherlands |
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Abstract: | On-chip sample preparation in self-contained microfluidic devices is a key element to realize simple, low-cost, yet reliable in vitro diagnostics that can be carried out at the point-of-care (POC) with minimal training requirements by unskilled users. To address this largely unmet POC medical need, we have developed an optimized polysaccharide matrix containing the reagents which substantially improves our fully printed POC CD4 counting chambers for the monitoring of HIV patients. The simply designed counting chambers allow for capillary-driven filling with unprocessed whole blood. We carefully tailored a gellan/trehalose matrix for deposition by inkjet printing, which preserves the viability of immunostains during a shelf life of at least 3 months and enables controlled antibody release for intense and homogeneous immunofluorescent cell staining throughout the complete 60 mm2 image area within 30 min. Excellent agreement between CD4 counts obtained from our fully printed CD4 counting chambers and the gold standard, flow cytometry, is demonstrated using samples both from healthy donors and HIV-infected patients.Gellan/trehalose layers were tailored to optimize on-chip storage and release of antibodies in a simple point-of-care CD4 counting chip with excellent agreement with standard methods. |
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