A novel phase shifting structured illumination microscopy

Veena Singh; Vishesh Dubey; Azeem Ahmad; Gyanendra Singh; D. S. Mehta

doi:10.1117/12.2212456

9 March 2016 A novel phase shifting structured illumination microscopy

Veena Singh, Vishesh Dubey, Azeem Ahmad, Gyanendra Singh, D. S. Mehta

Proceedings Volume 9713, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIII; 97131K (2016) https://doi.org/10.1117/12.2212456
Event: SPIE BiOS, 2016, San Francisco, California, United States

Abstract

This paper describes a new and novel phase shifting technique for qualitative as well as quantitative measurement in microscopy. We have developed a phase shifting device which is robust, inexpensive and involves no mechanical movement. In this method, phase shifting is implemented using LED array, beam splitters and defocused projection of Ronchi grating. The light from the LEDs are made incident on the beam splitters at spatially different locations. Due to variation in the geometrical distances of LEDs from the Ronchi grating and by sequentially illuminating the grating by switching on one LED at a time the phase shifted grating patterns are generated. The phase shifted structured patterns are projected onto the sample using microscopic objective lens. The phase shifted deformed patterns are recorded by a CCD camera. The initial alignment of the setup involves a simple procedure for the calibration for equal fringe width and intensity such that the phase shifted fringes are at equal phase difference. Three frame phase shifting algorithm is employed for the reconstruction of the phase map. The method described here is fully automated so that the phase shifted images are recorded just by switching of LEDs and has been used for the shape measurement of microscopic industrial objects. The analysis of the phase shifted images provides qualitative as well as quantitative information about the sample. Thus, the method is simple, robust and low cost compared to PZT devices commonly employed for phase shifting.

Conference Presentation

Citation Download Citation

Veena Singh, Vishesh Dubey, Azeem Ahmad, Gyanendra Singh, and D. S. Mehta "A novel phase shifting structured illumination microscopy", Proc. SPIE 9713, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIII, 97131K (9 March 2016); https://doi.org/10.1117/12.2212456

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