Magneto-Fluorescent Microbeads for Bacteria Detection Constructed from Superparamagnetic Fe₃O₄ Nanoparticles and...

The efficient and sensitive detection of pathogenic microorganisms in aqueous environments, such as water used in medical applications, drinking water, and cooling water of industrial plants, requires simple and fast methods suitable for multiplexed detection such as flow cytometry (FCM) with optically encoded carrier beads. For this purpose, we combine fluorescent Cd-free Ag-In-S ternary quantum dots (t-QDs) with fluorescence lifetimes (LTs) of several hundred nanoseconds and superparamagnetic Fe₃O₄ nanoparticles (SPIONs) with mesoporous CaCO₃ microbeads to a magneto-fluorescent bead platform that can be surface-functionalized with bioligands, such as antibodies. This inorganic bead platform enables immuno-magnetic separation, target enrichment, and target quantification with optical readout. The beads can be detected with steady-state and time-resolved fluorescence microscopy and flow cytometry (FCM). Moreover, they are suited for readout by time gated emission. In the following, the preparation of these magneto-fluorescent CaCO₃ beads, their spectroscopic and analytic characterization, and their conjugation with bacteria-specific antibodies are presented as well as proof-of-concept measurements with Legionella pneumophila including cell cultivation and plating experiments for bacteria quantification. Additionally, the possibility to discriminate between the long-lived emission of the LT-encoded capture and carrier CaCO₃ beads and the short-lived emission of the dye-stained bacteria with time-resolved fluorescence techniques and single wavelength excitation is demonstrated.