Double-Color MINFLUX Imaging of Microtubules and Vesicles

Area of Science:

• Cell Biology
• Biophysics
• Microscopy

Background:

• Microtubules and vesicles are vital for intracellular transport and signal transduction.
• Conventional microscopy lacks the resolution to detail their subcellular positional relationships.
• Understanding these dynamics requires advanced imaging techniques.

Purpose of the Study:

• To apply MINFLUX super-resolution microscopy for high-precision 2D and 3D imaging of microtubules and vesicles.
• To investigate the spatial relationships between microtubules and vesicles within the cellular cytoskeleton.
• To demonstrate MINFLUX's capability for resolving nanoscale interactions in live cells.

Main Methods:

• Utilized MINFLUX super-resolution microscopy for 2D and 3D imaging in BSC-1 cells.
• Achieved nanometer-scale localization precision for single molecules.
• Employed the detector channel ratio (DCR) method for distinguishing microtubule and vesicle signals.

Main Results:

• MINFLUX achieved ~4 nm precision in 2D and ~6 nm precision in 3D imaging.
• Successfully differentiated and assigned localizations to microtubules and vesicles.
• Visualized the precise spatial arrangement of microtubules and vesicles within the cytoskeletal network.

Conclusions:

• MINFLUX microscopy provides unprecedented spatial resolution for studying organelle and cytoskeletal interactions.
• The DCR method effectively separates signals from different cellular components in super-resolution imaging.
• This technique offers a powerful tool for dissecting the mechanisms of intracellular transport and signaling pathways.