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Expansion Microscopy of Bacillus subtilis.
Viola Middelhauve1, Jan Peter Siebrasse1, Ulrich Kubitscheck2
1Institute of Physical and Theoretical Chemistry, University of Bonn, Bonn, Germany.
Methods in Molecular Biology (Clifton, N.J.)
|November 29, 2022
Summary
Expansion microscopy offers super-resolved bacterial imaging using standard equipment. This protocol details visualizing Bacillus subtilis DivIVA-GFP and cell walls, highlighting protein and organism selection challenges.
Area of Science:
- Microscopy
- Cell Biology
- Biotechnology
Background:
- Expansion microscopy provides super-resolution imaging without specialized optical instruments.
- The technique utilizes conventional chemicals and laboratory equipment for sample preparation.
- Standard fluorescence microscopy is sufficient for imaging prepared specimens.
Purpose of the Study:
- To describe a detailed protocol for expansion microscopy of Bacillus subtilis.
- To demonstrate the visualization of DivIVA-GFP protein and cell wall.
- To emphasize challenges in selecting appropriate proteins and organisms for expansion microscopy.
Main Methods:
- Protocol for expansion microscopy applied to Bacillus subtilis.
- Expression of DivIVA-GFP fusion protein in Bacillus subtilis.
- Labeling of the bacterial cell wall using wheat germ agglutinin.
Main Results:
- Successful super-resolved imaging of Bacillus subtilis using expansion microscopy.
- Visualization of DivIVA-GFP localization within the bacterial cell.
- Demonstration of cell wall labeling for structural context.
Conclusions:
- Expansion microscopy is a viable method for high-resolution bacterial imaging with accessible tools.
- Careful selection of proteins and organisms is crucial for successful expansion microscopy protocols.
- The described protocol provides a foundation for further super-resolution studies in bacteria.
