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Related Experiment Videos

Time-resolved delayed luminescence image microscopy using an europium ion chelate complex

G Marriott1, M Heidecker, E P Diamandis

  • 1Department of Cell Biology, Max Planck Institute for Biochemistry, München, Germany.

Biophysical Journal
|September 1, 1994
PubMed
Summary

Time-resolved delayed luminescence imaging microscopy (TR-DLIM) offers high-contrast cell imaging. This technique utilizes a novel europium ion complex (SBMC) for enhanced visualization of biological molecules, even in living cells.

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Area of Science:

  • Cell biology
  • Microscopy
  • Biophysics

Background:

  • Delayed luminescence imaging microscopy (TR-DLIM) offers unique advantages for biological imaging.
  • Europium ion complexes provide high quantum yield and resistance to photobleaching and oxygen quenching.
  • Existing probes like eosin have limitations in sensitivity and environmental stability.

Purpose of the Study:

  • To describe improvements and extended applications of TR-DLIM in cell biology.
  • To introduce a novel streptavidin-based macromolecular complex (SBMC) for enhanced delayed luminescence imaging.
  • To demonstrate the capability of TR-DLIM for high-contrast imaging of labeled ligands in living and fixed cells.

Main Methods:

  • Utilizing europium ion complexed to a fluorescent chelating group for protein labeling.

Related Experiment Videos

  • Employing a streptavidin-based macromolecular complex (SBMC) for enhanced signal detection.
  • Exciting the fluorescent chelate at 340 nm and detecting europium ion emission at 612 nm.
  • Exploiting TR-DLIM's ability to reject scattered light, autofluorescence, and prompt fluorescence.

    • Main Results:

    • The SBMC complex exhibits a quantum yield orders of magnitude higher than eosin.
    • High-contrast images of biotin-labeled ligands were obtained, visible even to the naked eye.
    • Delayed luminescence imaging of living cells in aerated medium was achieved, complementing prompt fluorescence studies.
    • Double emission images of living amoeba cells were resolved, showing intracellular and surface-bound labels.
    • Environmental sensitivity of europium ion emission lifetime was observed in fixed cells.

    Conclusions:

    • TR-DLIM, enhanced by the SBMC probe, provides a powerful tool for high-contrast cell biology imaging.
    • The SBMC probe's properties overcome limitations of traditional probes, enabling imaging in challenging conditions.
    • The system allows for multiplexed imaging and environmental sensing through luminescence lifetime analysis.
    • The coupling of SBMC to streptavidin opens avenues for diverse immunocytochemical studies using biotin-based tracers.