Automatic full compensation of quantitative phase imaging in off-axis digital holographic microscopy

Area of Science:

• Optical microscopy
• Holography
• Image processing

Background:

• Quantitative phase measurements in off-axis digital holographic microscopy (DHM) are susceptible to perturbations.
• These perturbations, including curvature phase flaws from microscope objectives and reference wave tilt, degrade the accuracy of phase maps.
• Accurate phase measurements are crucial for analyzing microscopic samples.

Purpose of the Study:

• To develop and present an automatic method for fully compensating quantitative phase measurements in off-axis DHM.
• To eliminate the two main sources of phase error in off-axis DHM: microscope objective curvature and reference wave tilt.
• To improve the quality of recovered phase maps for both biological and nonbiological samples.

Main Methods:

• Utilized an optimized telecentric imaging system to avoid curvature phase flaws during hologram recording.
• Implemented automatic computation of a digital compensating reference wave to remove phase perturbations from reference wave tilt.
• Tested the developed method on both nonbiological and biological samples.

Main Results:

• Successfully demonstrated an automatic method for complete compensation of quantitative phase measurements in off-axis DHM.
• The method effectively removed phase errors originating from the microscope objective and reference wave tilt.
• Achieved significant improvement in the quality of recovered phase maps.

Conclusions:

• The presented automatic method offers a robust solution for accurate quantitative phase measurements in off-axis DHM.
• This technique enhances the reliability and quality of phase imaging for diverse microscopic applications.
• The method's effectiveness is validated across both biological and nonbiological sample analyses.