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qFlow Cytometry-Based Receptoromic Screening: A High-Throughput Quantification Approach Informing Biomarker Selection
Si Chen1, Jared Weddell1, Pavan Gupta2
1Bioengineering Department, University of Illinois at Urbana-Champaign, Urbana, IL, USA.
Methods in Molecular Biology (Clifton, N.J.)
|February 27, 2017
Summary
Quantitative flow cytometry (qFlow) offers precise, cell-by-cell biomarker quantification, overcoming limitations of traditional methods for improved medical diagnosis and nanosensor development.
Area of Science:
- Biomedical Engineering
- Molecular Biology
- Immunology
Background:
- Nanosensor development for medical diagnosis requires precise biomarker targeting.
- Immunohistochemistry (IHC) offers relative biomarker mapping but lacks cell-level detail and absolute quantification.
- Flow cytometry provides cell-by-cell analysis, and with calibration, enables quantitative biomarker measurement (qFlow cytometry).
Purpose of the Study:
- To outline the methodology for applying qFlow cytometry to quantify biomarkers.
- To focus on the angiogenic vascular endothelial growth factor receptor family as a target.
- To enable high-throughput quantification of biomarker expression for improved diagnostic strategies.
Main Methods:
- Antibody specificity testing and immunofluorescent cell labeling.
- Saturation analysis and fluorescent microsphere calibration for accurate quantitation.
- Quantitative analysis of both ensemble and cell-by-cell data.
Main Results:
- Established a robust qFlow cytometry protocol for biomarker detection.
- Enabled absolute quantification of biomarker concentrations on a cell-by-cell basis.
- Demonstrated high-throughput capability for biomarker expression profiling.
Conclusions:
- qFlow cytometry overcomes IHC limitations by providing absolute, cell-level biomarker quantification.
- This methodology is crucial for informed biomarker selection in nanosensor development.
- Enables precise measurement of biomarker expression for advanced medical diagnostics.
Keywords:
AngiogenesisBackground subtractionHeterogeneityImmuno-labelingMixture modelingPlatelet-Derived Growth Factor (PDGF)Quantitative flow cytometrySystems biologyVascular Endothelial Growth Factor (VEGF)qFlow cytometry
