Related Experiment Video
Updated: Aug 20, 2025
11:28
3D Orbital Tracking in a Modified Two-photon Microscope: An Application to the Tracking of Intracellular Vesicles
Published on: October 1, 2014
10.3K
Comprehensive Characterization of an "Off/On" Rhodol-Based Lysosomal Tracker for Orthogonal Cellular Analysis by
Carolina Chieffo1, Emiliano Altamura2, Layth Ben Trad1
1Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, ICBMS, UMR 5246), Université de Lyon, Université Claude Bernard Lyon 1, Bâtiment Edgar Lederer, 1 Rue Victor Grignard, 69622, Lyon, France.
Chembiochem : a European Journal of Chemical Biology
|November 24, 2022
Summary
New fluorescent lysosomal trackers were synthesized from fluorescein and rhodol. These novel xanthene-cyanamide dyes offer stable fluorescence in protic solvents, unlike traditional rhodamines, enabling clearer cellular imaging.
Area of Science:
- Organic Chemistry
- Biochemistry
- Cell Biology
Background:
- Rhodamine dyes are widely used fluorescent probes but can suffer from pH and solvent sensitivity.
- Electron-withdrawing groups (EWGs) are known to influence the photophysical properties of rhodamine derivatives.
- Developing stable, bright fluorescent trackers is crucial for advanced cellular imaging techniques.
Purpose of the Study:
- To synthesize novel fluorescent xanthene-cyanamide lysosomal trackers.
- To characterize the photophysical properties and stability of these new dyes.
- To evaluate their utility in cellular imaging applications.
Main Methods:
- Microwave-assisted synthesis of fluorescein and rhodol methyl esters.
- Characterization using NMR spectroscopy, X-ray analysis, and fluorimetry.
- Confocal microscopy imaging in the MC3T3-E1 osteoblastic cell line.
Main Results:
- Two novel fluorescent xanthene-cyanamide trackers emitting at ~525 nm were successfully prepared.
- Both "off" (nonfluorescent lactam) and "on" (fluorescent ring-opened amide) forms were characterized.
- Unlike rhodamines, the ring-opened fluorescein- and rhodol-cyanamides exhibit stability in protic solvents across a range of pH conditions.
- Effective differentiation of organelles and nuclei was achieved in cellular imaging.
Conclusions:
- Fluorescein- and rhodol-cyanamides represent a new class of stable lysosomal trackers.
- The introduction of cyanamide as an EWG enhances dye stability in protic media.
- These novel dyes show significant potential for advanced biological imaging and diagnostics.
