Immersion Mirau interferometry for label-free live cell imaging in an epi-illumination geometry

Oleksandra V. Lyulko; Gerhard Randers-Pehrson; David J. Brenner

doi:10.1117/12.855651

26 February 2010 Immersion Mirau interferometry for label-free live cell imaging in an epi-illumination geometry

Oleksandra V. Lyulko, Gerhard Randers-Pehrson, David J. Brenner

Proceedings Volume 7568, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VIII; 756825 (2010) https://doi.org/10.1117/12.855651
Event: SPIE BiOS, 2010, San Francisco, California, United States

Abstract

In cell biology studies it is often important to avoid the damaging effects caused by fluorescent stains or UV-light. Immersion Mirau Interferometry (IMI) is an epi-illumination label-free imaging technique developed at the Columbia University Radiological Research Accelerator Facility. It is based on the principles of phase-shifting interferometry (PSI) and represents a novel approach for interferometric imaging of living cells in medium. To accommodate the use of medium, a custom immersion Mirau interferometric attachment was designed and built in-house. The space between the reference mirror and the beam splitter is filled with liquid to ensure identical optical paths in the test and reference arms. The interferometer is mountable onto a microscope objective. The greatest limitation of standard PSI is the sensitivity to environmental vibrations, because it requires consecutive acquisition of several interferograms. We are developing Simultaneous Immersion Mirau Interferometry (SIMI), which facilitates simultaneous acquisition of all interferograms and eliminates the effects of vibration. Polarization optics, incorporated into the design, introduces a phase delay to one of the components of the test beam. This enables simultaneous creation and spatial separation of two interferograms, which are combined with the background image to reconstruct the intensity map of the specimen. Our results of imaging live and fixed cells with IMI and SIMI show that this system produces images of a quality that is sufficient to perform targeted cellular irradiation experiments.

Citation Download Citation

Oleksandra V. Lyulko, Gerhard Randers-Pehrson, and David J. Brenner "Immersion Mirau interferometry for label-free live cell imaging in an epi-illumination geometry", Proc. SPIE 7568, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VIII, 756825 (26 February 2010); https://doi.org/10.1117/12.855651

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