Common-path, single-shot phase-shifting digital holographic microscopy using a Ronchi ruling

Abstract

Phase-shifting digital holography is widely considered to be a groundbreaking method to quantitatively investigate the phase distribution of specimens, such as living cells. The main flaws of this method, however, are that the requirement for several sequential phase-shifted holograms eliminates the possibility of single-shot imaging and complex configurations would also increase the temporal noise. The present paper aims to validate a single-shot, common-path, phase-shifting digital holographic microscopy, employing a self-referencing geometry. A Ronchi ruling, located in the Fourier plane of a standard microscopic imaging system, produces multiple replicas of sample information in the image plane. The phase retrieval algorithm is performed by superposition of the sample-free portion of each replica with the object information, and requires at least three adjacent diffraction orders. To evaluate the performance of the proposed method, the phase distribution of silica microspheres as a test sample and the morphology of red blood cells as a biological specimen are determined. This configuration offers improved temporal stability in comparison with previously reported Mach-Zehnder interferometers, and may serve as an alternative for real-time surveying of nanometric and subnanometric fluctuations of living microscopic specimens.