SLM-based Fourier Differential Interference Contrast Microscopy

Abstract

Optical phase microscopy provides a view of objects that have minimal to no effect on the detected intensity of light that are unobservable by standard microscopy techniques. Since its inception just over 60 years ago that gave us a vision to an unseen world and earned Frits Zernike the Nobel prize in physics in 1953, phase microscopy has evolved to find various applications in biological cell imaging, crystallography, semiconductor failure analysis, and more. Two common and commercially available techniques are phase contrast and differential interference contrast (DIC). In phase contrast method, a large portion of the unscattered light that accounts for the majority of the light passing unaffected through a transparent medium is blocked to allow the scattered light due to the object to be observed with higher contrast. DIC is a self-referenced interferometer that transduces phase variation to intensity variation. While being established as fundamental tools in many scientific and engineering disciplines, the traditional implementation of these techniques lacks the ability to provide the means for quantitative and repeatable measurement without an extensive and cumbersome calibration. The rapidly growing fields in modern biology meteorology and nano-technology have emphasized the demand for a more robust and convenient quantitative phase microscopy. The recent emergence of modern optical devices such as high resolution programmable spatial light modulators (SLM) has enabled a multitude of research activities over the past decade to reinvent phase microscopy in unconventional ways. This work is concerned with an implementation of a DIC microscope containing a 4-f system at its core with a programmable SLM placed at the frequency plane of the imaging system that allows for employing Fourier pair transforms for wavefront manipulation. This configuration of microscope provides a convenient way to perform both wavefront shearing with quantifiable arbitrary shear amount and direction as well as phase stepping interferometry by programming the SLM with a series of numerically generated patterns and digitally capturing interferograms for each step which are then used to calculate the objects phase gradient map. Wavefront shearing is performed by generating a pattern for the SLM where two phase ramp patterns with opposite slopes are interleaved through a random selection process with uniform distribution in order to mimic the simultaneous presence of the ramps on the same plane. The theoretical treatment accompanied by simulations and experimental results and discussion are presented in this work.