Fully refractive telecentric f-theta microscope based on adaptive elements for 3D raster scanning of biological...

Area of Science:

• Optical microscopy
• Biomedical imaging
• Advanced optical systems

Background:

• Point-wise acquisition in microscopy (e.g., confocal, two-photon) enables sectioned imaging but requires 3D scanning.
• Traditional 3D scanning methods often use mechanical movement or reflective optics, leading to folded beam paths and potential aberrations.

Purpose of the Study:

• To introduce a fully refractive microscope capable of flexible three-dimensional scanning.
• To demonstrate aberration-free volumetric imaging with high resolution over a large field of view.

Main Methods:

• Utilized a combination of an adaptive lens, an adaptive prism, and a telecentric f-theta objective.
• Employed only transmissive optical components for a refractive setup.
• Performed 3D scanning over large ranges (X/Y=6300 μm, Z=480 μm).

Main Results:

• Achieved flexible 3D scanning with low scan-induced aberrations.
• Maintained uniform resolution across a large tuning range.
• Demonstrated volumetric measurements on zebrafish embryo fluorescence and pollen grains.

Conclusions:

• This refractive microscope represents a significant advancement for aberration-free volumetric imaging of 3D biological samples.
• The technology paves the way for smart microscopy solutions meeting demands in both lateral and axial dimensions without compromising image quality.
• Potential applications include imaging of embryos and organoids.