Rational design of fluorescent barcodes for suspension array through a simple simulation strategy

Abstract

Quantum dot (QD)-encoded microbeads as optical barcode with high fluorescence intensity and fluorescence uniformity, excellent stability and dispersity are greatly important for suspension array (SA). However, the size distribution of the microbeads mass-produced by the membrane emulsification method usually shows polydispersity, which leads to obstacles, imposing labour-intensive experimental iterations for the application of fluorescence-encoded microbeads as a distinguishable barcode. Herein, a simple simulation strategy based on a multicolor fluorescence model (MFM) was used to predict the influence of the microbeads’ size distribution on the barcode signals. The point L and S respectively represent the two end points of the barcode, and the line segment LS can be considered as a cluster of the QD-encoded microbeads (simulated barcode). Experimental clusters of fluorescent microbeads were found to be in good agreement with the simulated barcodes. This simple simulation strategy can effectively simplify the experimental iteration process because the fluorescence-encoded microbeads are not decoded by a flow cytometer. Moreover, when applied for the high-throughput ultrasensitive detection of three tumor markers (CEA, CA125 and CA199) in a single sample, these barcodes exhibit superior detection performance. Detection limits of 0.028 ± 0.001 ng mL⁻¹ for CEA, 1.5 ± 0.02 KU L⁻¹ for CA125 and 0.8 ± 0.1 KU L⁻¹ for CA199 are achieved, which meet the sensitivity criteria of tumor marker analysis. Therefore, this simple simulation strategy helps to overcome technical and economic obstacles for the widespread application of SA.