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Register a new userHighly stable common-path quantitative phase microscope for biomedical imaging
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High temporal stability is the primary requirement of any quantitative phase microscope (QPM) systems for the early stage detection of various human related diseases. The high temporal stability of the system provides accurate measurement of membrane fluctuations of the biological cells, which can be good indicator of various diseases. We developed a single element highly stable common-path QPM system to obtain temporally stable holograms of the biological specimens. With the proposed system, the temporal stability is obtained ~ 15 mrad without using any vibration isolation table. The capability of the proposed system is demonstrated on USAF resolution chart, polystyrene spheres (dia. 4.5 micron) and human red blood cells (RBCs). The membrane fluctuation of healthy human RBCs is further successfully measured and found to be equal to 63 nm. Contrary to its counterparts, present system offers energy efficient, cost effective and simple way of generating object and reference beam for the development of common-path QPM.
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Quantitative phase microscopy (QPM) has found significant applications in the field of biomedical imaging which works on the principle of interferometry. The theory behind achieving interference in QPM with conventional light sources such as white light and lasers is very well developed. Recently, the use of dynamic speckle illumination (DSI) in QPM has attracted attention due to its advantages over conventional light sources such as high spatial phase sensitivity, single shot, scalable field of view (FOV) and resolution. However, the understanding behind obtaining interference fringes in QPM with DSI has not been convincingly covered previously. This imposes a constraint on obtaining interference fringes in QPM using DSI and limits its widespread penetration in the field of biomedical imaging. The present article provides the basic understanding of DSI through both simulation and experiments that is essential to build interference optical microscopy systems such as QPM, digital holographic microscopy and optical coherence tomography. Using the developed theory of DSI we demonstrate its capabilities of using non-identical objective lenses in both arms of the interference microscopy without degrading the interference fringe contrast and providing the flexibility to use user-defined microscope objective lens. It is also demonstrated that the interference fringes are not washed out over a large range of optical path difference (OPD) between the object and the reference arm providing competitive edge over low temporal coherence light sources. The theory and explanation developed here would enable wider penetration of DSI based QPM for applications in biology and material sciences.
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