Assessing Membrane Fluidity and Visualizing Fluid Membrane Domains in Bacteria Using Fluorescent Membrane Dyes.
Michaela Wenzel1,2, Norbert O E Vischer1, Henrik Strahl3
1Bacterial Cell Biology, Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam, The Netherlands.
Bio-Protocol
|September 17, 2021
Summary
Bacterial membrane fluidity adapts to environments and impacts antibiotic effectiveness. New methods using fluorescent dyes laurdan and DiIC12 assess membrane fluidity and microdomains in bacteria like Bacillus subtilis.
Area of Science:
- Microbiology
- Biophysics
- Cell Biology
Background:
- Bacterial membrane fluidity is crucial for adaptation and antibiotic action.
- Membrane fluidity varies across the cell, forming specialized microdomains.
- Understanding these variations is key to bacterial cell organization and antibiotic mechanisms.
Purpose of the Study:
- To describe methods for assessing bacterial membrane fluidity and microdomain formation.
- To utilize fluorescent membrane dyes for evaluating membrane properties in living bacteria.
Main Methods:
- Employing two fluorescent membrane dyes, laurdan and DiIC12.
- Assessing membrane fluidity and microdomain formation in living bacteria, focusing on Bacillus subtilis.
- Adapting assays for other bacterial species like Staphylococcus aureus and Streptococcus pneumoniae.
Main Results:
- Demonstrated utility of laurdan and DiIC12 for assessing bacterial membrane fluidity.
- Provided adaptable methods for studying membrane dynamics in various bacterial species.
Conclusions:
- Laurdan and DiIC12 are effective tools for analyzing bacterial membrane fluidity and microdomains.
- These methods advance the understanding of bacterial membrane organization and antibiotic interactions.
Related Concept Videos
01:23Membrane Fluidity
162.2K
Cell membranes are composed of phospholipids, proteins, and carbohydrates loosely attached to one another through chemical interactions. Molecules are generally able to move about in the plane of the membrane, giving the membrane its flexible nature called fluidity. Two other features of the membrane contribute to membrane fluidity: the chemical structure of the phospholipids and the presence of cholesterol in the membrane.
162.2K
01:24Protein Diffusion in the Membrane
4.9K
Proteins show rotational as well as lateral diffusion across the membrane. The lateral diffusion of proteins was confirmed through the cell fusion experiment where mouse and human cells were fused, resulting in hybrid cells. When the human and mouse cells fused, the specific membrane proteins on human and mouse cells were marked with the red and green-fluorescent markers, respectively. Initially, the red and green fluorescence was located on the respective hemisphere of the cell. As time...
4.9K
01:19Protein Dynamics in Living Cells
2.4K
Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
2.4K