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      Multi-Contrast Imaging and Digital Refocusing on a Mobile Microscope with a Domed LED Array

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          Abstract

          We demonstrate the design and application of an add-on device for improving the diagnostic and research capabilities of CellScope—a low-cost, smartphone-based point-of-care microscope. We replace the single LED illumination of the original CellScope with a programmable domed LED array. By leveraging recent advances in computational illumination, this new device enables simultaneous multi-contrast imaging with brightfield, darkfield, and phase imaging modes. Further, we scan through illumination angles to capture lightfield datasets, which can be used to recover 3D intensity and phase images without any hardware changes. This digital refocusing procedure can be used for either 3D imaging or software-only focus correction, reducing the need for precise mechanical focusing during field experiments. All acquisition and processing is performed on the mobile phone and controlled through a smartphone application, making the computational microscope compact and portable. Using multiple samples and different objective magnifications, we demonstrate that the performance of our device is comparable to that of a commercial microscope. This unique device platform extends the field imaging capabilities of CellScope, opening up new clinical and research possibilities.

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          Most cited references29

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          Wide-field, high-resolution Fourier ptychographic microscopy

          In this article, we report an imaging method, termed Fourier ptychographic microscopy (FPM), which iteratively stitches together a number of variably illuminated, low-resolution intensity images in Fourier space to produce a wide-field, high-resolution complex sample image. By adopting a wavefront correction strategy, the FPM method can also correct for aberrations and digitally extend a microscope’s depth-of-focus beyond the physical limitations of its optics. As a demonstration, we built a microscope prototype with a resolution of 0.78 μm, a field-of-view of ~120 mm2, and a resolution-invariant depth-of-focus of 0.3 mm (characterized at 632 nm). Gigapixel colour images of histology slides verify FPM’s successful operation. The reported imaging procedure transforms the general challenge of high-throughput, high-resolution microscopy from one that is coupled to the physical limitations of the system’s optics to one that is solvable through computation.
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            Mobile Phone Based Clinical Microscopy for Global Health Applications

            Light microscopy provides a simple, cost-effective, and vital method for the diagnosis and screening of hematologic and infectious diseases. In many regions of the world, however, the required equipment is either unavailable or insufficiently portable, and operators may not possess adequate training to make full use of the images obtained. Counterintuitively, these same regions are often well served by mobile phone networks, suggesting the possibility of leveraging portable, camera-enabled mobile phones for diagnostic imaging and telemedicine. Toward this end we have built a mobile phone-mounted light microscope and demonstrated its potential for clinical use by imaging P. falciparum-infected and sickle red blood cells in brightfield and M. tuberculosis-infected sputum samples in fluorescence with LED excitation. In all cases resolution exceeded that necessary to detect blood cell and microorganism morphology, and with the tuberculosis samples we took further advantage of the digitized images to demonstrate automated bacillus counting via image analysis software. We expect such a telemedicine system for global healthcare via mobile phone – offering inexpensive brightfield and fluorescence microscopy integrated with automated image analysis – to provide an important tool for disease diagnosis and screening, particularly in the developing world and rural areas where laboratory facilities are scarce but mobile phone infrastructure is extensive.
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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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                Author and article information

                Contributors
                Role: Academic Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                2015
                13 May 2015
                : 10
                : 5
                : e0124938
                Affiliations
                [1 ]Graduate Group in Applied Science and Technology, University of California, Berkeley, CA 94720, USA
                [2 ]Department of Electrical Engineering and Computer Sciences, University of California, Berkeley, CA 94720, USA
                [3 ]Department of Bioengineering, University of California, Berkeley, CA 94720, USA
                Glasgow University, UNITED KINGDOM
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: LW DAF ZFP MVD LT N. Switz. Performed the experiments: ZFP JJR MVD N. Switz LT LW. Analyzed the data: ZFP JJR LW LT N. Sadras AVG DAF N. Switz HSP. Wrote the paper: ZFP MVD LT N. Switz DAF LW.

                Article
                PONE-D-14-57810
                10.1371/journal.pone.0124938
                4430423
                25969980
                a0be1f6e-85ae-4e3b-b7c9-1e18083bcc02
                Copyright @ 2015

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

                History
                : 24 December 2014
                : 19 March 2015
                Page count
                Figures: 5, Tables: 0, Pages: 13
                Funding
                This work was supported by USAID Cooperative Agreements AID-OAA-A-13-00002 and AID-OAA-A-12-00011, part of USAID's Higher Education Solutions Network, and by the Gordon and Betty Moore Foundation's Data-Driven Discovery Initiative through grant GBMF4562 to Laura Waller.
                Categories
                Research Article
                Custom metadata
                All relevant data are available from Figshare at the following URL: http://figshare.com/articles/Computational_CellScope_LED_Dome_Design_Files/1354054.

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