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Area of Science:

  • Microbiology
  • Biotechnology
  • Analytical Chemistry

Background:

  • Distinguishing between dormant and active bacterial spores is crucial for public health and safety.
  • Existing methods, like the Schaeffer-Fulton method, are often destructive, time-consuming, and labor-intensive.
  • There is a need for rapid, non-destructive techniques for bacterial spore analysis.

Purpose of the Study:

  • To develop a simple, quick, and non-destructive method for differentiating dormant and germinated bacterial spores.
  • To utilize lanthanide-beta-diketonate complexes for direct single-spore visualization.

Main Methods:

  • Employing lanthanide-beta-diketonate complexes for instantaneous staining of bacterial spores.
  • Observing differential staining patterns between dormant (nonfluorescent core) and germinated (fluorescent core) spores.
  • Utilizing a USB microscope for low-cost imaging of spores in various matrices, including milk.

Main Results:

  • The lanthanide complexes successfully stained dormant and germinated spores from five bacterial species, including *Bacillus*, *Clostridium*, and *Clostridioides*.
  • Dormant spores showed a nonfluorescent core, while germinated spores exhibited a brightly fluorescent core.
  • The method is rapid, requires minimal sample processing, and allows for live imaging due to low complex toxicity.

Conclusions:

  • Lanthanide-beta-diketonate complexes provide an effective, non-destructive method for distinguishing bacterial spore states.
  • This technique offers a valuable, low-cost tool for studying bacterial spores in diverse settings, enhancing biosecurity and food safety.
  • The versatility in complex formulation allows for tailored spore visualization.