Abstract
In recent years, the development of new image analysis approaches has highlighted the possibility of recovering super-resolution information from short sequences of wide-field images. Our recently developed method, SRRF (Super-Resolution Radial Fluctuations), enables long-term live-cell imaging beyond the resolution limit without specialized hardware. Here, we present eSRRF (enhanced-SRRF), a significant improvement over our initial method, enhancing image fidelity to the underlying structure and resolution. Especially, eSRRF uses automated data-driven parameter optimization, including an estimation of the number of frames necessary for optimal reconstruction. We demonstrate the improved fidelity of the images reconstructed with eSRRF and highlight its versatility and ease of use over a wide range of microscopy techniques and biological systems. We also extend eSRRF to 3D super-resolution microscopy by combining it with multi-focus microscopy (MFM), obtaining volumetric super-resolution imaging of live cells with acquisition speed of ∼ 1 volume/second.
Competing Interest Statement
The authors have declared no competing interest.
Footnotes
Romain F. Laine LaineBioImaging
Hannah S. Heil: Hannah_SuperRes
Simao Coelho: simaopc
Jonathon Nixon-Abell: AbellJonny
Aki Stubb: akistub
Gautier Follain: @Follain_Ga
Siãn Culley: SuperResoluSian
Guillaume Jacquemet: guijacquemet
Bassam Hajj: Bassam_A_HAJJ
Christophe Leterrier: christlet
Ricardo Henriques: HenriquesLab
Figure 3, Methods, Supplementary Movies