Magneto-Fluorescent Microbeads for Bacteria Detection Constructed from Superparamagnetic Fe3O4 Nanoparticles and AIS/ZnS Quantum Dots

Anal Chem. 2019 Oct 15;91(20):12661-12669. doi: 10.1021/acs.analchem.9b01812. Epub 2019 Sep 30.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Immobilized / chemistry
  • Antibodies, Immobilized / immunology
  • Calcium Carbonate / chemical synthesis
  • Calcium Carbonate / chemistry
  • Coloring Agents / chemistry
  • Ferrosoferric Oxide / chemistry
  • Flow Cytometry / methods
  • Legionella pneumophila / immunology
  • Legionella pneumophila / isolation & purification*
  • Magnetite Nanoparticles / chemistry*
  • Microscopy, Fluorescence / methods*
  • Microspheres
  • Quantum Dots / chemistry*
  • Silver / chemistry
  • Sulfides / chemistry
  • Sulfur / chemistry
  • Zinc Compounds / chemistry

Substances

  • Antibodies, Immobilized
  • Coloring Agents
  • Magnetite Nanoparticles
  • Sulfides
  • Zinc Compounds
  • Silver
  • Sulfur
  • Calcium Carbonate
  • zinc sulfide
  • Ferrosoferric Oxide