The new live imagers MitoMM1/2 for mitochondrial visualization
Miwa Maeda1, Mayu Suzuki1, Shigeo Takashima2
1Department of Chemistry and Biomolecular Science, Faculty of Engineering, Gifu University, 1-1 Yanagido, Gifu, 501-1193, Japan.
Abstract:
Mitochondria are eukaryotic organelles that consist of outer and inner bilayer membranes with a positive potential (H+) in the intermembrane space. This organelle plays an important role in ATP production and apoptosis. To observe the mitochondria in living cells, several fluorescent dyes (such as MitoTracker® [a standard mitochondrial imager] or rhodamine 123) have been developed. However, these reagents are unstable and exhibit a wide range of emission spectra, thereby hampering double staining results. Using recombinant DNA techniques, green or red fluorescent protein (GFP or RFP)-tagged proteins are now available for multi-color labeling of mitochondria. Here, we have discussed the development of the novel mitochondrial live imagers MitoMM1/2, derivatives of ATTO565; furthermore, MitoMM1/2 are sensitive to the membrane potential, resistant to detergents, and the fluorescence of MitoMM1/2 does not overlap with green fluorescence.
Insights
Researchers developed new fluorescent dyes, MitoMM1/2, for imaging mitochondria in living cells. These novel dyes are stable, sensitive to membrane potential, and do not interfere with green fluorescent proteins, enabling better multi-color imaging.
Area of Science:
- Cell Biology
- Biochemistry
- Molecular Imaging
Background:
- Mitochondria are vital organelles involved in ATP production and apoptosis.
- Current fluorescent dyes for mitochondria imaging, like MitoTracker®, have limitations including instability and broad emission spectra, hindering multi-color labeling.
- Genetically encoded fluorescent proteins (GFP, RFP) offer multi-color labeling but may not always be optimal for specific imaging needs.
Purpose of the Study:
- To develop novel, stable, and sensitive fluorescent dyes for live-cell mitochondrial imaging.
- To overcome the limitations of existing mitochondrial imaging reagents, particularly for multi-color applications.
- To create mitochondrial imagers with fluorescence properties that do not overlap with common green fluorescent proteins.
Main Methods:
- Development of novel mitochondrial live imagers, MitoMM1/2, as derivatives of ATTO565.
- Characterization of MitoMM1/2 for sensitivity to mitochondrial membrane potential.
- Assessment of MitoMM1/2 resistance to detergents and spectral properties for multi-color compatibility.
Main Results:
- MitoMM1/2 were successfully synthesized as novel mitochondrial live imagers.
- These dyes demonstrated sensitivity to mitochondrial membrane potential.
- MitoMM1/2 exhibited detergent resistance and fluorescence spectra that do not overlap with green fluorescence, facilitating double staining.
- The developed dyes offer improved stability and spectral characteristics compared to traditional reagents.
Conclusions:
- MitoMM1/2 represent advanced fluorescent probes for live-cell mitochondrial imaging.
- These novel imagers enhance the capability for multi-color mitochondrial studies by avoiding spectral overlap with green fluorescent proteins.
- MitoMM1/2 provide a valuable tool for researchers studying mitochondrial dynamics, function, and apoptosis in living cells.
