Fluorescence-labeling in bacterial imaging

Session

Poster Session

Authors

Sophie Neumann (1, 2), Christina Wichmann (1, 3, 4), Aurélie Jost (2), Patrick Then (2), Christian Eggeling (1, 3)

Affiliations

1. Institute for Applied Optics and Biophysics, Friedrich Schiller University Jena
2. Microverse Imaging Center, Cluster of Excellence Balance of the Microverse, Friedrich Schiller University Jena
3. Leibniz-Institute of Photonic Technologies e.V. Jena
4. Cluster of Excellence Balance of the Microverse, Friedrich Schiller University Jena

Keywords

bacterial imaging, spectral unmixing, Staphylococcus cohnii, Escherichia coli

Abstract text

We show new insight into the application of fluorescence labeling techniques for the visualization and discrimination of Gram-positive and Gram-negative bacteria using confocal laser scanning microscopy (CLSM).

The Microverse Imaging Center (MIC) in Jena, Germany, is an advanced light microscopy imaging facility supporting other scientists with their imaging experiments. Especially biologists benefit from the services offered by imaging facilities.

Assisting microbiological imaging projects with imaging methods employing fluorescent labels instead of genetically modifying the microbes has many advantages. It is not only a faster method but also allows for the imaging to take place in laboratories not having a safety clearance for genetically modified organisms (GMOs).

The study compares the performance of several widely used fluorophores for fluorescent imaging of bacterial samples. All dyes were used to stain Staphylococcus cohnii as a representative gram-positive bacterial strain and Escherichia coli as a representative gram-negative bacterial strain.

Performing spectrally resolved confocal imaging over time as well as spectral unmixing, all dyes were tested in both S. cohnii and E. coli to characterize the staining behavior of these dyes in both bacterial strains.

Our findings offer practical insights for researchers employing fluorescence labeling in microbiological studies. Additionally, they contribute to the refinement of confocal imaging techniques for bacterial discrimination. This work not only helps us as an imaging facility support our users. It also enhances our understanding of bacterial cell biology and paves the way for improved diagnostic and therapeutic strategies targeting specific bacterial types.