An assembly-regulated SNAP-tag fluorogenic probe for long-term super-resolution imaging of mitochondrial dynamics

Area of Science:

• Cell Biology
• Microscopy
• Biochemistry

Background:

• Super-resolution fluorescence microscopy offers insights into mitochondrial dynamics.
• Limitations in probe photostability and chemstability hinder long-term imaging of mitochondria.

Purpose of the Study:

• To develop a novel probe for enhanced super-resolution imaging of mitochondrial dynamics.
• To overcome limitations of existing probes for long-term live-cell imaging.

Main Methods:

• Synthesis of a novel SNAP-tag probe, AN-BG, derived from 4-azetidinyl-naphthliamide.
• Utilizing the probe's fluorogenicity and photostability for super-resolution microscopy.
• Labeling mitochondrial inner membrane proteins via SNAP tags for stable mitochondrial visualization.

Main Results:

• AN-BG exhibits excellent fluorogenicity and photostability.
• The probe's planar structure suppresses intramolecular charge transfer, enhancing brightness and stability.
• Stable mitochondrial labeling and long-term super-resolution imaging of dynamics were achieved, independent of mitochondrial membrane potential.

Conclusions:

• AN-BG is a suitable probe for long-term super-resolution imaging of mitochondrial dynamics.
• The probe design overcomes previous challenges in mitochondrial live-cell imaging.
• This work advances the study of cellular processes using advanced microscopy techniques.