13 March 2014 High-resolution quantitative imaging of subcellular morphology and cell refractometry in a liquid environment via endogenous mechanism
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Abstract
Biological cells are composed primarily of water; and as such are challenging to image without staining since the induced intensity modulation of transmitted or reflected light is typically insufficient to permit acceptable contrast for optical imaging. This issue may be resolved with the aid of exogenous contrast agents, but this often has a deleterious effect on the cell and precludes in vivo imaging. A unique approach to this problem is afforded by the phase contrast microscope in which optical-path differences in transmitted light is exploited as a contrast mechanism for qualitative imaging. In recent years however, several quantitative phase imaging techniques have been developed which allow for diffraction limited endogenous-contrast imaging with excellent temporal resolution. We hereby present a laser scanning technique for quantitative phase imaging which achieves sub-diffraction limited resolution at the expense of temporal resolution. This instrument is based on a stabilized fiber interfometer which is incorporated into a near-field scanning optical microscope (NSOM) for tri-modal imaging. Our latest results will focus on modifications made to this system to facilitate imaging in a liquid environment. A simple approach for achieving stable shear-force feedback operation in a liquid will be presented. Acquired high resolution images of white blood cells revealed detailed sub-cellular features. Images of fibroblast cells in air and in a liquid environment confirm the efficacy of the feedback operation in a liquid. Moreover, we demonstrate cell refractometry capability without the need for ad hoc modifications. These results clearly highlight the unique potential of this instrument for the study of living cells.
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Kert Edward, Faramarz Farahi, "High-resolution quantitative imaging of subcellular morphology and cell refractometry in a liquid environment via endogenous mechanism", Proc. SPIE 9038, Medical Imaging 2014: Biomedical Applications in Molecular, Structural, and Functional Imaging, 90380K (13 March 2014); doi: 10.1117/12.2044407; https://doi.org/10.1117/12.2044407
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