OPTiM: Optical projection tomography integrated microscope using open-source hardware and software.

Area of Science:

• Biomedical Imaging
• Microscopy Techniques
• Optical Engineering

Background:

• Optical Projection Tomography (OPT) is a powerful 3D imaging technique.
• Implementing OPT typically requires specialized, often expensive, equipment.
• Adapting existing microscopy platforms for advanced techniques like OPT can enhance accessibility.

Purpose of the Study:

• To describe the development and implementation of an open-source OPT plate.
• To integrate OPT capabilities into a commercial wide-field inverted microscope.
• To demonstrate multiple methods for performing OPT with varying magnification and resolution.

Main Methods:

• An OPT plate with tilt adjustment and stepper motor was designed and fabricated.
• Three distinct OPT acquisition methods were detailed: direct OPT, external optical-relay OPT, and remote focal scanning OPT.
• All methods utilized the microscope's existing incoherent light source, preserving its standard functionality.

Main Results:

• The OPT plate successfully enabled 3D tomographic imaging on a standard inverted microscope.
• Different OPT configurations achieved varying magnifications and spatial resolutions, with one method reaching ~1.6 μm.
• The system demonstrated viability through successful acquisitions on in vivo zebrafish embryos.

Conclusions:

• The developed open-source OPT plate provides a versatile and accessible solution for high-resolution 3D imaging.
• This implementation broadens the applicability of OPT to researchers with commercial inverted microscopes.
• The modular design and multiple imaging strategies offer flexibility for diverse biological imaging applications.