High numerical aperture holographic microscopy reconstruction with extended z range

TL;DR

This paper introduces a holographic microscopy reconstruction method compatible with high NA objectives up to 1.4, enabling extended z-range imaging with near-optimal resolution through calibration and phase mask techniques.

Contribution

The method combines phase mask application and exact kernel propagation to achieve high-NA holographic reconstruction over an extended z-range, validated experimentally.

Findings

Achieved near-optimal resolution over ±50 μm z-range.

Validated with high NA=1.4 microscope objective.

Effective calibration method for extended depth imaging.

Abstract

An holographic microscopy reconstruction method compatible with high numerical aperture microscope objective (MO) up to NA=1.4 is proposed. After off axis and reference field curvature corrections, and after selection of the +1 grating order holographic image, a phase mask that transforms the optical elements of the holographic setup into an afocal device is applied in the camera plane. The reconstruction is then made by the angular spectrum method. The field is first propagated in the image half space from the camera to the afocal image of the MO optimal plane (plane for which MO has been designed) by using a quadratic kernel. The field is then propagated from the MO optimal plane to the object with the exact kernel. Calibration of the reconstruction is made by imaging a calibrated object like an USAF resolution target for different positions along $z$. Once the calibration is done, the reconstruction can be made with an object located in any plane $z$. The reconstruction method has been validated experimentally with an USAF target imaged with a NA=1.4 microscope objective. Near-optimal resolution is obtained over an extended range ($\pm 50~\mu$m) of $z$ locations.