Abstract
This paper proposes an optimized implementation of the double-exposure method with emphasis on the uniformity and minimization of the residual phase imperfections in cross-referenced holographic microscopy (CRHM). The quantitative phase images are restored from single-shot cross-referenced holograms, which are separated in the Fourier space and processed to eliminate effects caused by imperfections of the optical path and sample background. CRHM is implemented in a microscope configuration supplemented by a Sagnac interference module providing splitting and shearing of the sample and reference waves. Utilization of the averaging process, which enhances precision of quantitative phase image (QPI) reconstruction, applicable in the methods with a replicated field of view is also presented. The high temporal stability of CRHM is verified in calibration measurements and its application potential demonstrated by a quantitative restoration of the phase resolution target and imaging of biological samples including cheek and sperm cells.
Highlights
Quantitative phase imaging (QPI) is a widely used method for providing information about spatial variations in the optical thickness of weakly absorbing objects that enables to measure the refractive index[1] and the dry mass density distribution of individual cells[2]
The single-shot hologram reconstruction maintaining the high temporal stability is feasible in a common-path setup, in which interfering waves share nearly the same optical path
In this publication the optimal use of the double-exposure method often used in the cross-referenced holographic microscopy (CRHM) for aberration elimination is presented
Summary
Quantitative phase imaging (QPI) is a widely used method for providing information about spatial variations in the optical thickness of weakly absorbing objects that enables to measure the refractive index[1] and the dry mass density distribution of individual cells[2]. In methods with a replicated field of view, one of the beams can be filtered in the Fourier plane to create an isolated reference beam with reduced information about the object[9,10]. This approach enables imaging of the dense samples but requires a precise alignment of added optical components. CRHM works in a stable common-path geometry allowing a single-shot quantitative restoration of phase images, while enhancement of the imaging performance is achieved by processing the cross-referenced holograms
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