Related Experiment Video
Updated: May 6, 2026
11:27
Single-Molecule Förster Resonance Energy Transfer Methods for Real-Time Investigation of the Holliday Junction Resolution by GEN1
Published on: September 18, 2019
10.0K
A Fluorogenic Reaction for Monitoring Cross-Coupling with Turn-On Ratio Greater than a Hundred Thousand
Rachel V Czerwinski1, Benjamin A Brewster1, Sunil P Upadhyay1
1Department of Chemistry, University of Wisconsin-Madison, 1101 University Ave., Madison, Wisconsin 53705, United States.
Chemical & Biomedical Imaging
|February 27, 2026
Summary
Researchers developed a new fluorogenic reaction design achieving unprecedented 207,000 turn-on ratios. This breakthrough enables sensitive detection in chemical and biological imaging by minimizing reactant fluorescence.
Area of Science:
- Chemical Sciences
- Molecular Biology
- Biophysical Chemistry
Background:
- Fluorogenic reactions are vital tools in molecular sciences for detection and imaging.
- Existing fluorogenic reactions suffer from high reactant fluorescence, limiting their sensitivity and application range.
Purpose of the Study:
- To design a novel fluorogenic reaction with exceptionally high turn-on ratios.
- To overcome the limitations imposed by residual reactant fluorescence in sensitive detection methods.
Main Methods:
- Developed a design scheme based on a condensation reaction between electron-rich and electron-deficient molecules.
- Utilized Suzuki cross-coupling to synthesize a dicyanomethylenedihydrofuran (DCDHF) product.
- Characterized the photophysical properties, including absorption and Stokes shift, of reactants and products.
Main Results:
- Achieved turn-on ratios up to 207,000, significantly exceeding previous limits.
- Synthesized a DCDHF product with absorption ~1 eV redder than reactants and a large Stokes shift (~0.5 eV).
- Demonstrated negligible reactant excitation, enabling high reactant concentrations without signal interference.
Conclusions:
- The developed fluorogenic reaction design enables highly sensitive detection due to minimal reactant fluorescence.
- This approach broadens the applicability of fluorogenic reactions in high-concentration assays and advanced imaging.
- The reaction's performance was validated in both bulk conditions and microdroplets.
