A fluorogenic probe for β-galactosidase activity imaging in living cells.

Area of Science:

• Biochemistry
• Cell Biology
• Molecular Imaging

Background:

• Beta-galactosidase is a key enzyme in cellular processes.
• Accurate imaging of beta-galactosidase activity is crucial for biological research.
• Existing probes may have limitations in signal duration or background noise.

Purpose of the Study:

• To develop a novel, cell-permeable fluorescence turn-on probe for beta-galactosidase.
• To enable sensitive and long-lasting imaging of beta-galactosidase activity in living cells.
• To overcome limitations of existing imaging techniques.

Main Methods:

• Synthesis and characterization of the AcGQCy7 probe.
• Cellular uptake and hydrolysis studies in beta-galactosidase-expressing cells.
• Confocal microscopy to assess fluorescence signal localization and duration.
• Evaluation of background fluorescence in culture media.

Main Results:

• AcGQCy7 demonstrated cell permeability and efficient internalization by beta-galactosidase-expressing cells.
• Hydrolysis of AcGQCy7 resulted in a highly fluorescent molecule.
• The fluorescent signal was retained in cellular mitochondria for several days.
• A strong, beta-galactosidase-dependent intracellular signal with low background fluorescence was achieved.

Conclusions:

• AcGQCy7 is an effective fluorescence turn-on probe for imaging beta-galactosidase activity.
• The probe provides a long-lasting and sensitive intracellular fluorescent signal.
• AcGQCy7 is suitable for long-term monitoring of beta-galactosidase expression in live cells.