Interferometric microscopy has grown into a very potent tool for quantitative phase imaging of biological samples. Among the interfermetric methods, microscopy by digital holography is one of the most effective techniques, especially for studying dynamics of cells. Imaging of cell fluctuations requires digital holographic setups with high temporal stability. Common path setups in which the object and the reference beams encounter the same set of optical elements provide better temporal stability compared to two-beam setups. Here, we present a compact, easy-to-implement, common path digital holographic microscope based on Sagnac interferometer geometry. The microscope is implemented using a diode laser module employing a CCD array or a webcam sensor to record holograms. The system was tested for three-dimensional imaging capability, numerical focusing ability, and temporal stability. Sub-nanometer temporal stability without external vibration isolation components was obtained in both cases. The higher temporal stability makes the microscope compatible to image cell fluctuations, which is demonstrated by imaging the oscillation of the cell membrane of human red blood cells.

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Digital holographic microscopy is the state of the art quantitative phase imaging of micro-objects including living cells. It is an ideal tool to image and quantify cell thickness profiles with nanometer thickness resolution. Digital holographic techniques usually are implemented using a two-beam setup that may be bulky and may not be field portable. Self-referencing techniques provide compact geometry but suffer from a reduction of the field of view. Here, we discuss the development of a wavefront division digital holographic microscope providing the full field of view with a compact system. The proposed approach uses a wavefront division module consisting of two lenses. The developed microscope is tested experimentally by measuring the physical and mechanical properties of red blood cells.

Quantitative three-dimensional (3-D) imaging of living cells provides important information about the cell morphology and its time variation. Off-axis, digital holographic interference microscopy is an ideal tool for 3-D imaging, parameter extraction, and classification of living cells. Two-beam digital holographic microscopes, which are usually employed, provide high-quality 3-D images of micro-objects, albeit with lower temporal stability. Common-path digital holographic geometries, in which the reference beam is derived from the object beam, provide higher temporal stability along with high-quality 3-D images. Self-referencing geometry is the simplest of the common-path techniques, in which a portion of the object beam itself acts as the reference, leading to compact setups using fewer optical elements. However, it has reduced field of view, and the reference may contain object information. Here, we describe the development of a common-path digital holographic microscope, employing a shearing plate and converting one of the beams into a separate reference by employing a pin-hole. The setup is as compact as self-referencing geometry, while providing field of view as wide as that of a two-beam microscope. The microscope is tested by imaging and quantifying the morphology and dynamics of human erythrocytes.