Optical sectioning robotic microscopy for everyone: the structured illumination microscope with the OpenFlexure...

Area of Science:

• Microscopy
• Optical Engineering
• Biotechnology

Background:

• Traditional microscopy techniques often lack optical sectioning capabilities, leading to out-of-focus blur.
• Confocal microscopy offers optical sectioning but can be expensive and complex.
• 3D printing offers a pathway to creating affordable and customizable scientific instrumentation.

Purpose of the Study:

• To develop a low-cost, 3D-printed structured illumination microscope (SIM) with optical sectioning capabilities.
• To integrate precise, open-source 3D-printable stages for enhanced imaging control.
• To achieve optical sectioning performance comparable to confocal microscopy.

Main Methods:

• Utilized 3D-printed components from the OpenFlexure family for precise stage actuation (nanometer precision).
• Implemented structured illumination microscopy by projecting grid patterns and acquiring phase-shifted images.
• Employed simple mathematical post-processing to reconstruct optically sectioned images.
• Integrated Raspberry Pi for automated and remote operation via wireless local area network.

Main Results:

• Achieved optical sectioning strength of a few microns, comparable to confocal microscopes.
• Demonstrated precise sample positioning and grid actuation using 3D-printed flexure-based stages.
• Successfully automated microscope operation and enabled remote control.

Conclusions:

• The 3D-printed SIM provides an accessible and cost-effective alternative for achieving high-quality optical sectioning.
• Open-source 3D-printable components enable the construction of advanced microscopy systems.
• The developed system has potential applications in various biological and material science imaging fields.