Parallel on-axis holographic phase microscopy of biological cells and unicellular microorganism dynamics.

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2010-05-20

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Abstract

We apply a wide-field quantitative phase microscopy technique based on parallel two-step phase-shifting on-axis interferometry to visualize live biological cells and microorganism dynamics. The parallel on-axis holographic approach is more efficient with camera spatial bandwidth consumption compared to previous off-axis approaches and thus can capture finer sample spatial details, given a limited spatial bandwidth of a specific digital camera. Additionally, due to the parallel acquisition mechanism, the approach is suitable for visualizing rapid dynamic processes, permitting an interferometric acquisition rate equal to the camera frame rate. The method is demonstrated experimentally through phase microscopy of neurons and unicellular microorganisms.

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Wax

Adam P. Wax

Professor of Biomedical Engineering

Dr. Wax's research interests include optical spectroscopy for early cancer detection, novel microscopy and interferometry techniques.

The study of intact, living cells with optical spectroscopy offers the opportunity to observe cellular structure, organization and dynamics in a way that is not possible with traditional methods. We have developed a set of novel spectroscopic techniques for measuring spatial, temporal and refractive structure on sub-hertz and sub-wavelength scales based on using low-coherence interferometry (LCI) to detect scattered light. We have applied these techniques in different types of cell biology experiments. In one experiment, LCI measurements of the angular pattern of backscattered light are used to determine non-invasively the structure of sub-cellular organelles in cell monolayers, and the components of epithelial tissue from freshly excised rat esophagus. This work has potential as a diagnostic method for early cancer detection. In another experiment, LCI phase measurements are used to examine volume changes of epithelial cells in a monolayer in response to environmental osmolarity changes. Although cell volume changes have been measured previously, this work demonstrates for the first time the volume of just a few cells (2 or 3) tracked continuously and in situ.


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