β-Galactosidase- and Photo-Activatable Fluorescent Probes for Protein Labeling and Super-Resolution STED Microscopy in Living Cells.
Taukeer A KhanStefan StoldtMariano L BossiVladimir N BelovStefan W HellPublished in: Molecules (Basel, Switzerland) (2024)
We report on the synthesis of two fluorescent probes which can be activated by β-Galactosidase (β-Gal) enzymes and/or light. The probes contained 2-nitro-4-oxybenzyl and 3-nitro-4-oxybenzyl fragments, with β-Gal residues linked to C-4. We performed the enzymatic and photoactivation of the probes in a cuvette and compared them, prior to the labeling of Vimentin-Halo fusion protein in live cells with overexpressed β-galactosidase. The dye fluorescence afforded the observation of enzyme activity by means of confocal and super-resolution optical microscopy based on stimulated emission depletion (STED). The tracing of enzymatic activity with the retention of activated fluorescent products inside cells was combined with super-resolution imaging as a tool for use in biomedicine and life science.
Keyphrases
- living cells
- single molecule
- fluorescent probe
- high resolution
- induced apoptosis
- cell cycle arrest
- high speed
- optical coherence tomography
- endoplasmic reticulum stress
- public health
- hydrogen peroxide
- fluorescence imaging
- quantum dots
- protein protein
- single cell
- oxidative stress
- photodynamic therapy
- cell proliferation
- highly efficient
- binding protein