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Updated: May 13, 2026
07:44
Application of Membrane and Cell Wall Selective Fluorescent Dyes for Live-Cell Imaging of Filamentous Fungi
Published on: November 28, 2019
Turn-on, fluorescent nuclear stains with live cell compatibility
Demar R G Pitter1, Jens Wigenius, Adrienne S Brown
1Department of Chemistry, University of Miami, 1301 Memorial Drive, Coral Gables, Florida 33146, USA.
Organic Letters
|March 7, 2013
Summary
New green and yellow fluorescent probes bind to DNA, offering bright and clear imaging for live cells. These probes are superior alternatives to traditional nuclear stains, showing significant emission enhancement upon DNA binding.
Area of Science:
- Molecular Biology
- Biochemistry
- Organic Chemistry
Background:
- Traditional nuclear stains like Hoechst 33342 and DAPI have limitations, including UV excitation requirements and lower fluorescence.
- There is a need for advanced fluorescent probes with improved brightness, photostability, and compatibility with live-cell imaging techniques.
Purpose of the Study:
- To develop novel green and yellow fluorescent probes for DNA visualization.
- To evaluate the probes' performance in terms of brightness, photostability, and live-cell compatibility.
- To elucidate the DNA-binding mechanisms of the novel probes.
Main Methods:
- Synthesis of novel green and yellow fluorescent DNA-binding probes.
- Optical spectroscopy (absorption, emission, fluorescence lifetime) and time-dependent density-functional theory (TD-DFT) calculations to investigate electronic structure.
- Linear dichroism measurements to determine DNA binding modes (groove binding vs. intercalation).
- Live-cell imaging experiments to assess membrane permeability and cellular compatibility.
Main Results:
- The synthesized probes exhibit excellent brightness and high on/off ratios.
- Probes are membrane permeable and compatible with live-cell imaging.
- DNA binding results in significant emission enhancements (27- to 75-fold).
- Linear dichroism confirms distinct DNA binding modes: one probe acts as a groove binder, the other as an intercalator.
- Probes are optimally matched to 405 nm and 514 nm laser lines, offering an alternative to UV-excited stains.
Conclusions:
- The novel green and yellow fluorescent probes represent a significant advancement in DNA visualization tools.
- Their superior optical properties and live-cell compatibility make them valuable for various biological imaging applications.
- The probes provide mechanistic insights into DNA-dye interactions, distinguishing between groove binding and intercalation.
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Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
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