Ultrabright fluorescent microsphere and its novel application for improving the sensitivity of immunochromatographic...

Fluorescent microsphere (FM) is widely used as probe in immunochromatographic assay (ICA). However, the performance of conventional FM is limited because of the aggregation-caused quenching effect. Herein, we compared a kind of conventional FM (DMFFM, loading DMF) with novel aggregation-induced emission FM (AIEFM, loading TCBPE). The fluorescence intensity of DMFFM initially increased and then decreased as the concentrations of the loading DMF increased. The fluorescence intensity of AIEFM increased as the concentrations of the loading TCBPE increased and retained a high value. AIEFM was compared with two commercial FMs purchased from Ocean (OFM) and Merk (MFM). The maximum fluorescence intensity and relative quantum yield of AIEFM was approximately 5 and 4.5 times higher than those of two commercial FMs. We used the novel AIEFM as a probe to improve the sensitivity of ICA. When Escherichia coli O157:H7 was detected as the target, the limit of detection of ICA based on AIEFM, OFM and MFM were 3.98 × 10³ CFU/mL, 4.48 × 10⁴ and 2.78 × 10⁴ CFU/mL, respectively. The ICA of AIEFM had 11 and 7 times improvement in sensitivity compared with that of OFM and MFM. Our results could be used as a basis for novel probes in practical ICA applications.