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Related Concept Videos

Immunofluorescence Microscopy01:12

Immunofluorescence Microscopy

A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.
The...

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Tracking Synthetic Adhesins on Bacterial Surfaces with Immunofluorescence Microscopy.

Sofía Fraile1, Jaime Fernández de Córdoba2, Víctor de Lorenzo3

  • 1Department of Systems Biology, Centro Nacional de Biotecnología-CSIC, 28049, Madrid, Spain.

Methods in Molecular Biology (Clifton, N.J.)
|July 4, 2026
PubMed
Summary

Researchers developed a method to visualize synthetic adhesins on bacteria using super-resolution microscopy. This technique allows for tracking engineered proteins on bacterial surfaces for applications in biocatalysis and drug delivery.

Keywords:
Bacterial cell surfaceImmunofluorescenceNanobodiesSTED microscopySurface displaySynthetic adhesins

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Biotechnology

Background:

  • Programmable bacterial adhesion is crucial for advanced applications like biocatalysis and drug delivery.
  • Synthetic adhesins, hybrid proteins engineered for specific functions, are key to achieving this bacterial surface modification.
  • Visualizing these adhesins is essential for understanding and optimizing their function.

Purpose of the Study:

  • To describe a detailed protocol for visualizing synthetic adhesins on the surface of Escherichia coli and Pseudomonas putida.
  • To demonstrate the utility of indirect immunofluorescence combined with Super-Resolution Microscopy (SRM) for this purpose.

Main Methods:

  • Construction and expression of synthetic adhesins in bacterial hosts.
  • Indirect immunofluorescence staining targeting specific adhesin components.
  • Super-Resolution Microscopy (SRM), specifically STimulated Emission Depletion (STED) microscopy, for high-resolution imaging.
  • Image acquisition and spatial analysis of adhesin localization on the bacterial cell surface.

Main Results:

  • Successful visualization of synthetic adhesins on the surface of E. coli and P. putida.
  • High-resolution spatial tracking of adhesin distribution using STED microscopy.
  • Validation of the protocol for monitoring expression and localization of engineered bacterial surface proteins.

Conclusions:

  • The described protocol provides a robust method for visualizing synthetic adhesins on bacteria.
  • This technique enables detailed spatial analysis of engineered bacterial surface proteins, crucial for developing new biotechnological tools.
  • Super-resolution microscopy is a powerful approach for studying bacterial surface display systems.