1. Homogeneous agglutination assay based on micro-chip sheathless flow cytometry
- Author
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Zengshuai Ma, Xie Shuai, Xiongying Ye, Pan Zhang, Zhang Shuai, and Yinuo Cheng
- Subjects
Fluid Flow and Transfer Processes ,Detection limit ,Analyte ,Materials science ,Chromatography ,biology ,medicine.diagnostic_test ,Molecular biophysics ,Microfluidics ,Biomedical Engineering ,Analytical chemistry ,Condensed Matter Physics ,Chip ,Flow cytometry ,Agglutination (biology) ,Colloid and Surface Chemistry ,biology.protein ,medicine ,General Materials Science ,Fabrication and Laboratory Methods ,Bovine serum albumin - Abstract
Homogeneous assays possess important advantages that no washing or physical separation is required, contributing to robust protocols and easy implementation which ensures potential point-of-care applications. Optimizing the detection strategy to reduce the number of reagents used and simplify the detection device is desirable. A method of homogeneous bead-agglutination assay based on micro-chip sheathless flow cytometry has been developed. The detection processes include mixing the capture-probe conjugated beads with an analyte containing sample, followed by flowing the reaction mixtures through the micro-chip sheathless flow cytometric device. The analyte concentrations were detected by counting the proportion of monomers in the reaction mixtures. Streptavidin-coated magnetic beads and biotinylated bovine serum albumin (bBSA) were used as a model system to verify the method, and detection limits of 0.15 pM and 1.5 pM for bBSA were achieved, using commercial Calibur and the developed micro-chip sheathless flow cytometric device, respectively. The setup of the micro-chip sheathless flow cytometric device is significantly simple; meanwhile, the system maintains relatively high sensitivity, which mainly benefits from the application of forward scattering to distinguish aggregates from monomers. The micro-chip sheathless flow cytometric device for bead agglutination detection provides us with a promising method for versatile immunoassays on microfluidic platforms.
- Published
- 2015