Axial coding in full-field microscopy using three-dimensional structured illumination implemented with no moving...

Area of Science:

• Microscopy and Imaging Science
• Optical Physics
• Nanotechnology

Background:

• Structured illumination microscopy (SIM) enhances lateral resolution.
• Axial localization in microscopy remains a challenge for 3D imaging.
• Combining axial and lateral super-resolution techniques is desirable.

Purpose of the Study:

• To develop a simple optical setup for simultaneous axial and lateral structured illumination.
• To achieve high-accuracy axial localization of nanoscale objects.
• To maintain lateral resolution enhancement when combined with axial localization.

Main Methods:

• Utilizing a single objective lens for structured illumination.
• Acquiring a minimum of six full-field images without moving the sample or objective.
• Implementing a novel optical configuration for combined axial and lateral illumination.

Main Results:

• Accurate axial localization of 100 nm fluorescent beads with 50 nm accuracy in a 1.76-microm layer.
• Demonstrated combination of axial localization with classical lateral SIM.
• Maintained a factor of 2 lateral resolution enhancement.

Conclusions:

• The presented optical setup offers a straightforward method for simultaneous 3D super-resolution imaging.
• The technique significantly improves axial localization accuracy in fluorescence microscopy.
• This approach is compatible with existing lateral SIM methods, enhancing 3D imaging capabilities.