Holographic microscopy reconstruction in both object and image half-spaces with an undistorted three-dimensional grid

Area of Science:

• Optics and Photonics
• Microscopy Techniques
• Digital Holography

Background:

• Holographic microscopy offers 3D imaging capabilities.
• Accurate reconstruction is crucial for quantitative analysis.
• Existing methods may suffer from reconstruction grid distortions.

Purpose of the Study:

• To develop a holographic microscopy reconstruction method.
• To achieve reconstructions with an undistorted grid (orthogonal axes, constant pixel pitch).
• To validate the method's performance across a range of object positions.

Main Methods:

• Hologram propagation in the object half-space.
• Utilizing a reconstruction numerical phase mask and reference phase curvature.
• Calibration considering the microscope objective (MO) as an afocal device.

Main Results:

• The proposed method yields reconstructions with an undistorted grid.
• Validated using a USAF target imaged with a ×60 MO.
• Successful reconstructions achieved for longitudinal positions from -75 to +75 μm.

Conclusions:

• The developed method provides accurate holographic microscopy reconstructions.
• The technique ensures geometric fidelity in the reconstructed volume.
• Reconstruction can be equivalently interpreted in object or image space.