Obtaining super-resolved images at the mesoscale through super-resolution radial fluctuations

Area of Science:

• Optical microscopy
• Cell biology
• Biophysics

Background:

• Current super-resolution imaging techniques have limited fields of view (FOV), potentially biasing results and hindering comprehensive analysis.
• Manual selection of regions of interest (ROIs) in microscopy can introduce bias, and stitching small ROIs can cause artifacts.

Purpose of the Study:

• To achieve accurate super-resolution images across a large FOV (4.4 × 3.0 mm).
• To overcome the limitations of small FOV in current super-resolution microscopy.
• To enable unbiased analysis of cellular structures and their large-scale interactions.

Main Methods:

• Application of super-resolution radial fluctuations (SRRF) processing.
• Utilizing the Mesolens, which combines low magnification with high numerical aperture.
• Combining SRRF with Mesolens for enhanced imaging.

Main Results:

• Achieved super-resolved images with a resolution of 446.3 ± 10.9 nm.
• Demonstrated a ~1.6-fold improvement in spatial resolution over a large FOV (4.4 × 3.0 mm).
• Maintained minimal error and consistent structural agreement in the large FOV images.

Conclusions:

• A simple method for obtaining accurate super-resolution images over a large FOV has been developed.
• This technique allows for simultaneous understanding of subcellular structures and their large-scale interactions.
• Enables comprehensive analysis of biological samples without compromising resolution or introducing bias.