Joint tagging assisted fluctuation nanoscopy enables fast high-density super-resolution imaging

Area of Science:

• Biophysics
• Optical Microscopy
• Nanotechnology

Background:

• Investigating high-density subcellular structures with high fidelity is challenging in fluctuation-based optical nanoscopy.
• Existing methods struggle to achieve both high resolution and high labeling density simultaneously.

Purpose of the Study:

• To develop a novel strategy for fast, high-density nanoscopic imaging and tracking of subcellular structures.
• To enhance the capabilities of super-resolution radial fluctuation (SRRF) microscopy for complex biological samples.

Main Methods:

• Employed a joint tagging (JT) strategy combined with super-resolution radial fluctuation (SRRF) microscopy.
• Utilized spectrally distinguishable quantum dots for labeling in fixed cell experiments.
• Applied 3-channel JT for live-cell tracking of lipid clusters.

Main Results:

• Achieved high-density super-resolution imaging of microtubules with ~62 nm resolution by combining spectral channels.
• Successfully tracked dynamic motions of high-density lipid clusters in live cells below the optical diffraction limit.
• Demonstrated high-fidelity reconstruction by decreasing labeling density in each spectral channel.

Conclusions:

• The joint tagging assisted SRRF technique enables high-density, high-fidelity super-resolution imaging and tracking.
• This method overcomes limitations in visualizing dense subcellular structures in both fixed and live biological systems.
• Offers a powerful tool for studying dynamic cellular processes at the nanoscale.