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Handheld Fluorescence Microscopy based Flow Analyzer

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Abstract

Fluorescence microscopy has the intrinsic advantages of favourable contrast characteristics and high degree of specificity. Consequently, it has been a mainstay in modern biological inquiry and clinical diagnostics. Despite its reliable nature, fluorescence based clinical microscopy and diagnostics is a manual, labour intensive and time consuming procedure. The article outlines a cost-effective, high throughput alternative to conventional fluorescence imaging techniques. With system level integration of custom-designed microfluidics and optics, we demonstrate fluorescence microscopy based imaging flow analyzer. Using this system we have imaged more than 2900 FITC labeled fluorescent beads per minute. This demonstrates high-throughput characteristics of our flow analyzer in comparison to conventional fluorescence microscopy. The issue of motion blur at high flow rates limits the achievable throughput in image based flow analyzers. Here we address the issue by computationally deblurring the images and show that this restores the morphological features otherwise affected by motion blur. By further optimizing concentration of the sample solution and flow speeds, along with imaging multiple channels simultaneously, the system is capable of providing throughput of about 480 beads per second.

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Acknowledgments

Authors gratefully acknowledge the funding from Department of Electronics and Information Technology (DeitY), and Department of Biotechnology (DBT) for supporting Innovative Young Biotechnologist Award (IYBA-2013) Project.

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Authors and Affiliations

  1. Optics & Microfluidics Instrumentation Lab, Department of Instrumentation and Applied Physics, Indian Institute of Science, Bangalore, 560012, India

    Manish Saxena, Nitin Jayakumar & Sai Siva Gorthi

  2. Indian Space Research Organization, Ahmedabad, 380015, India

    Manish Saxena

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  1. Manish Saxena
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  2. Nitin Jayakumar
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  3. Sai Siva Gorthi
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Correspondence to Nitin Jayakumar.

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Saxena, M., Jayakumar, N. & Gorthi, S.S. Handheld Fluorescence Microscopy based Flow Analyzer. J Fluoresc 26, 631–638 (2016). https://doi.org/10.1007/s10895-015-1749-0

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  • DOI: https://doi.org/10.1007/s10895-015-1749-0

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