Multi-beam spatially multiplexed interference microscopy for phase objects examination

Piotr Zdańkowski , Vicente Mico Mico , Jose Angel Picazo-Bueno , Krzysztof Patorski , Maciej Trusiak

Abstract

Multi-beam Spatially Multiplexed Interferometric Microscopy (MB-SMIM) is presented as the implementation of a robust, fast and accurate single-shot phase-estimation algorithm with a simple, low-cost and stable way to convert a bright-field microscope into a holographic one using partially-coherent illumination. Ronchi diffraction grating is used to generate two-beam and multi-beam interference. To ensure effectiveness of the process object field should be divided into analyzed specimen area (1/3) and clear reference area (other 2/3). Using partially coherent light source one is able to produce pure three-beam interference of -1 diffraction order of specimen area, and 0/+1 diffraction orders of both clear reference areas. Opposite sign orders create doubled spatial frequency term overlapping with the regular SMIM interferogram. Phase demodulation based on the single-frame two-dimensional Hilbert-Huang transform or regular multi-frame phase-shifting follows. Single-frame processing is more beneficial for increased spatial frequency term when doubled number of fringes “samples” the same phase variation as in regular SMIM interferogram. Adding (or subtracting) two three-beam interferograms phase shifted by π allows one to accurately isolate doubled (or single) spatial frequency fringe pattern. Altogether, MB-SMIM adds quantitative phase imaging capability to a commercially available non-holographic microscope with improved phase reconstruction: noise reduction, spatial frequency doubling and phase change disambiguation capabilities. Proposed method enables common-path optical generation of doubled spatial frequency fringes only by slight object field manipulation – no mechanical doubling of incident angle is needed. The technique has been validated using a 20X/0.46NA objective in Olympus BX-60 upright microscope for prostate cancer cells phase imaging.
Publication typeTranslation of publication
Translator Piotr Zdańkowski (FM / IMPh)
Piotr Zdańkowski,,
- The Institute of Micromechanics and Photonics
, Vicente Mico Mico - Universitat de Valencia
Vicente Mico Mico,,
-
, Jose Angel Picazo-Bueno - Universitat de Valencia
Jose Angel Picazo-Bueno,,
-
, Krzysztof Patorski (FM / IMPh)
Krzysztof Patorski,,
- The Institute of Micromechanics and Photonics
, Maciej Trusiak (FM / IMPh)
Maciej Trusiak,,
- The Institute of Micromechanics and Photonics
Corporate authorInstytut Mikromechaniki i Fotoniki, Wydział Mechatroniki, PW
Pages107490J-1 - 107490J-7
Publication size in sheets0.3
Book Creath Katherine, Burke Jan, Morris Michael B. North, Davies Angela D. (eds.): Proc. SPIE 10749, Interferometry XIX, 2018
Keywords in Englishquantitative phase imaging, interference microscopy, dual reference interferometry, biological phase imaging
DOIDOI:10.1117/12.2323607
URL https://www.spiedigitallibrary.org/conference-proceedings-of-spie/10749/107490J/Multi-beam-spatially-multiplexed-interference-microscopy-for-phase-objects-examination/10.1117/12.2323607.short?SSO=1
Languageen angielski
Score (nominal)0
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