ZOLA-3D allows flexible 3D localization microscopy over an adjustable axial range

Area of Science:

• Biophysics
• Microscopy
• Optical Imaging

Background:

• Single molecule localization microscopy (SMLM) enables 3D super-resolution imaging without scanning.
• Challenges exist in achieving optimal resolution over extended axial ranges in SMLM.

Purpose of the Study:

• To present Zernike Optimized Localization Approach in 3D (ZOLA-3D), a computational and optical solution.
• To achieve optimal resolution over a tunable axial range for 3D super-resolution microscopy.

Main Methods:

• Development of ZOLA-3D, integrating optical and computational strategies.
• Leveraging axial variations of point spread functions (PSFs) for improved depth imaging.

Main Results:

• Demonstrated ZOLA-3D's capability for 3D super-resolution imaging.
• Achieved optimal resolution over a tunable axial range.
• Successfully imaged mitochondria, nuclear pores, and microtubules up to ~5 μm deep within cells.

Conclusions:

• ZOLA-3D provides an easy-to-use solution for deep 3D super-resolution imaging.
• The method enhances resolution over extended axial ranges, overcoming previous limitations.
• Enables visualization of subcellular structures in entire nuclei or cells at unprecedented depths.