High-fidelity 3D live-cell nanoscopy through data-driven enhanced super-resolution radial fluctuation

authors

• Laine Romain F
• Heil Hannah S
• Coelho Simao
• Nixon-Abell Jonathon
• Jimenez Angélique
• Wiesner Theresa
• Martínez Damián
• Galgani Tommaso
• Régnier Louise
• Stubb Aki
• Follain Gautier
• Webster Samantha
• Goyette Jesse
• Dauphin Aurelien
• Salles Audrey
• Culley Siân
• Jacquemet Guillaume
• Hajj Bassam
• Leterrier Christophe
• Henriques Ricardo

keywords

• Image processing
• Single-molecule biophysics
• Super-resolution microscopy

abstract

Live-cell super-resolution microscopy enables the imaging of biological structure dynamics below the diffraction limit. Here we present enhanced super-resolution radial fluctuations (eSRRF), substantially improving image fidelity and resolution compared to the original SRRF method. eSRRF incorporates automated parameter optimization based on the data itself, giving insight into the trade-off between resolution and fidelity. We demonstrate eSRRF across a range of imaging modalities and biological systems. Notably, we extend eSRRF to three dimensions by combining it with multifocus microscopy. This realizes live-cell volumetric super-resolution imaging with an acquisition speed of ~1 volume per second. eSRRF provides an accessible super-resolution approach, maximizing information extraction across varied experimental conditions while minimizing artifacts. Its optimal parameter prediction strategy is generalizable, moving toward unbiased and optimized analyses in super-resolution microscopy.

more information