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      Profiling individual human red blood cells using common-path diffraction optical tomography


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          Due to its strong correlation with the pathophysiology of many diseases, information about human red blood cells (RBCs) has a crucial function in hematology. Therefore, measuring and understanding the morphological, chemical, and mechanical properties of individual RBCs is a key to understanding the pathophysiology of a number of diseases in hematology, as well as to opening up new possibilities for diagnosing diseases in their early stages. In this study, we present the simultaneous and quantitative measurement of the morphological, chemical, and mechanical parameters of individual RBCs employing optical holographic microtomography. In addition, it is demonstrated that the correlation analyses of these RBC parameters provide unique information for distinguishing and understanding diseases.

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          Refractive index maps and membrane dynamics of human red blood cells parasitized by Plasmodium falciparum.

          Parasitization by malaria-inducing Plasmodium falciparum leads to structural, biochemical, and mechanical modifications to the host red blood cells (RBCs). To study these modifications, we investigate two intrinsic indicators: the refractive index and membrane fluctuations in P. falciparum-invaded human RBCs (Pf-RBCs). We report experimental connections between these intrinsic indicators and pathological states. By employing two noninvasive optical techniques, tomographic phase microscopy and diffraction phase microscopy, we extract three-dimensional maps of refractive index and nanoscale cell membrane fluctuations in isolated RBCs. Our systematic experiments cover all intraerythrocytic stages of parasite development under physiological and febrile temperatures. These findings offer potential, and sufficiently general, avenues for identifying, through cell membrane dynamics, pathological states that cause or accompany human diseases.
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            Optical imaging of cell mass and growth dynamics.

            Using novel interferometric quantitative phase microscopy methods, we demonstrate that the surface integral of the optical phase associated with live cells is invariant to cell water content. Thus, we provide an entirely noninvasive method to measure the nonaqueous content or "dry mass" of living cells. Given the extremely high stability of the interferometric microscope and the femtogram sensitivity of the method to changes in cellular dry mass, this new technique is not only ideal for quantifying cell growth but also reveals spatially resolved cellular and subcellular dynamics of living cells over many decades in a temporal scale. Specifically, we present quantitative histograms of individual cell mass characterizing the hypertrophic effect of high glucose in a mesangial cell model. In addition, we show that in an epithelial cell model observed for long periods of time, the mean squared displacement data reveal specific information about cellular and subcellular dynamics at various characteristic length and time scales. Overall, this study shows that interferometeric quantitative phase microscopy represents a noninvasive optical assay for monitoring cell growth, characterizing cellular motility, and investigating the subcellular motions of living cells.
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              Diffraction phase microscopy for quantifying cell structure and dynamics.

              We have developed diffraction phase microscopy as a new technique for quantitative phase imaging of biological structures. The method combines the principles of common path interferometry and single-shot phase imaging and is characterized by subnanometer path-length stability and millisecond-scale acquisition time. The potential of the technique for quantifying nanoscale motions in live cells is demonstrated by experiments on red blood cells.

                Author and article information

                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group
                17 October 2014
                : 4
                : 6659
                [1 ]Department of Physics, Korea Advanced Institute of Science and Technology , Daejeon 305–701, Republic of Korea
                [2 ]Department of Laboratory Medicine, University of Ulsan, College of Medicine and Asan Medical Center , Seoul 138–736, Republic of Korea
                Author notes

                These authors contributed equally to this work.


                Current address: Photonics Group, Department of Physics, Imperial College London, London SW7 2AZ, United Kingdom.


                Current address: Seegene Medical Foundation, Seoul 133–847, Republic of Korea.

                Copyright © 2014, Macmillan Publishers Limited. All rights reserved

                This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder in order to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/4.0/

                : 24 June 2014
                : 29 September 2014



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