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qFlow Cytometry-Based Receptoromic Screening: A High-Throughput Quantification Approach Informing Biomarker Selection and Nanosensor Development

Chen, Si and Weddell, Jared and Gupta, Pavan and Conard, Grace and Parkin, James and Imoukhuede, Princess I. (2017) qFlow Cytometry-Based Receptoromic Screening: A High-Throughput Quantification Approach Informing Biomarker Selection and Nanosensor Development. In: Biomedical Nanotechnology. Methods in Molecular Biology. No.1570. Springer , New York, NY, pp. 117-138. ISBN 978-1-4939-6838-1.

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Nanosensor-based detection of biomarkers can improve medical diagnosis; however, a critical factor in nanosensor development is deciding which biomarker to target, as most diseases present several biomarkers. Biomarker-targeting decisions can be informed via an understanding of biomarker expression. Currently, immunohistochemistry (IHC) is the accepted standard for profiling biomarker expression. While IHC provides a relative mapping of biomarker expression, it does not provide cell-by-cell readouts of biomarker expression or absolute biomarker quantification. Flow cytometry overcomes both these IHC challenges by offering biomarker expression on a cell-by-cell basis, and when combined with calibration standards, providing quantitation of biomarker concentrations: this is known as qFlow cytometry. Here, we outline the key components for applying qFlow cytometry to detect biomarkers within the angiogenic vascular endothelial growth factor receptor family. The key aspects of the qFlow cytometry methodology include: antibody specificity testing, immunofluorescent cell labeling, saturation analysis, fluorescent microsphere calibration, and quantitative analysis of both ensemble and cell-by-cell data. Together, these methods enable high-throughput quantification of biomarker expression.

Item Type:Book Section
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Additional Information:© 2017 Springer Science+Business Media LLC.
Subject Keywords:Quantitative flow cytometry – qFlow cytometry – Immuno-labeling – Systems biology – Vascular Endothelial Growth Factor (VEGF) – Platelet-Derived Growth Factor (PDGF) – Angiogenesis – Background subtraction – Mixture modeling – Heterogeneity
Series Name:Methods in Molecular Biology
Issue or Number:1570
Record Number:CaltechAUTHORS:20170306-090340482
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:74759
Deposited By: Tony Diaz
Deposited On:06 Mar 2017 17:12
Last Modified:03 Oct 2019 16:42

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