The widespread adoption of bead-based multiplexed bioassays requires the ability to easily synthesize encoded microspheres and conjugate analytes of interest to their ***,we present a simple method(MRBLEs 2.0)for the ...
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The widespread adoption of bead-based multiplexed bioassays requires the ability to easily synthesize encoded microspheres and conjugate analytes of interest to their ***,we present a simple method(MRBLEs 2.0)for the efficient high-throughput generation of microspheres with ratiometric barcode lanthanide encoding(MRBLEs)that bear functional groups for downstream surface *** production in MRBLEs 2.0 relies on the manual mixing of lanthanide/polymer mixtures(each of which comprises a unique spectral code)followed by droplet generation using single-layer,parallel flow-focusing devices and the off-chip batch polymerization of droplets into *** streamline downstream analyte coupling,MRBLEs 2.0 crosslinks copolymers bearing functional groups on the bead surface during bead *** the MRBLEs 2.0 pipeline,we generate monodisperse MRBLEs containing 48 distinct well-resolved spectral codes with high throughput(>150,000/min and can be boosted to 450,000/min).We further demonstrate the efficient conjugation of oligonucleotides and entire proteins to carboxyl MRBLEs and of biotin to amino ***,we show that MRBLEs can also be magnetized via the simultaneous incorporation of magnetic nanoparticles with only a minor decrease in the potential code *** the advantages of dramatically simplified device fabrication,elimination of the need for custom-made equipment,and the ability to produce spectrally and magnetically encoded beads with direct surface functionalization with high throughput,MRBLEs 2.0 can be directly applied by many labs towards a wide variety of downstream assays,from basic biology to diagnostics and other translational research.
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