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Updated: Jun 6, 2025
09:45
Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
Published on: February 9, 2012
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Harnessing Cyanine-like Properties to Develop Bright Fluorogenic Probes Based on Viscosity-Sensitive Molecular
Blaise Dumat1, Carolina Chieffo1
1Laboratoire des biomolécules, LBM, Département de chimie, École normale supérieure, PSL University, Sorbonne Université, CNRS, 75005, Paris, France.
Chemistry (Weinheim an Der Bergstrasse, Germany)
|December 1, 2024
Summary
Researchers developed brighter fluorescent molecular rotors (FMRs) by incorporating a julolidine group. These new FMRs enable enhanced bioimaging, including a lysosomal pH sensor and brighter HaloTag probes for live-cell protein imaging.
Area of Science:
- Chemical Biology
- Biophysical Chemistry
- Molecular Imaging
Background:
- Dipolar fluorescent molecular rotors (FMRs) are sensitive to microenvironmental viscosity and polarity, useful for bioimaging and biomolecule labeling.
- Previous FMRs showed potential for live-cell protein imaging via HaloTag, but moderate absorption limited fluorescence brightness.
Purpose of the Study:
- To synthesize and characterize novel hemicyanine-based FMRs with enhanced photophysical properties, specifically increased molar absorption coefficients.
- To investigate the impact of a strong julolidine electron-donating group on combining intense absorption with viscosity sensitivity.
- To develop new fluorescent probes for bioimaging applications, including a lysosomal pH sensor and brighter HaloTag ligands.
Main Methods:
- Synthesis of three model hemicyanines on a styrylindolenium scaffold.
- Detailed photophysical characterization in solvents of varying polarity and viscosity.
- Application of developed fluorogens for lysosomal pH sensing and HaloTag-mediated live-cell imaging of membrane proteins.
Main Results:
- The incorporation of a strong julolidine electron-donating group successfully combined intense cyanine-like absorption with high viscosity sensitivity.
- Developed fluorogens exhibited significantly improved brightness compared to previous FMRs.
- Demonstrated utility in creating a functional lysosomal pH sensor and two bright, cell-impermeant HaloTag ligands for membrane protein imaging.
Conclusions:
- A bright fluorogenic scaffold based on a simple hemicyanine structure was developed, overcoming the brightness limitations of traditional FMRs.
- This new scaffold enables the creation of advanced fluorescent probes and sensors with efficient photophysical properties for diverse bioimaging applications.
- The findings pave the way for novel tools in live-cell imaging, diagnostics, and sensing.
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