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Optical Recording of Cellular Zinc Dynamics with Zinc-Finger-Based Biosensors

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Zinc Finger Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1867))

Abstract

In addition to serving as an essential structural component, zinc is also involved in intracellular and intercellular signaling pathways to impact a number of cellular functions. Genetically encoded zinc sensors that are specifically targeted to various subcellular compartments (ER, mitochondria, nucleus, plasma membrane, and vesicles) have been proven to provide accurate and sensitive visualization and quantification of zinc. Here we describe the methods to utilize both ratiometric and intensiometric genetically encoded zinc sensors designed based on zinc fingers for imaging and quantification of cellular free, labile zinc concentrations, [Zn2+]free. This chapter explains in detail how to quantify [Zn2+]free in live cells as well as how to monitor zinc influx in INS-1 cells stimulated with high glucose.

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Authors and Affiliations

  1. Department of Biological Sciences, University of Denver, Denver, CO, USA

    Dylan H. Fudge, Ray Black & Yan Qin

Authors
  1. Dylan H. Fudge
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  2. Ray Black
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  3. Yan Qin
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Corresponding author

Correspondence to Yan Qin .

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Editors and Affiliations

  1. Shanghai Institute for Advanced Immunochemical Studies, ShanghaiTech University, Shanghai, China

    Jia Liu

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Fudge, D.H., Black, R., Qin, Y. (2018). Optical Recording of Cellular Zinc Dynamics with Zinc-Finger-Based Biosensors. In: Liu, J. (eds) Zinc Finger Proteins. Methods in Molecular Biology, vol 1867. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8799-3_8

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  • DOI: https://doi.org/10.1007/978-1-4939-8799-3_8

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8798-6

  • Online ISBN: 978-1-4939-8799-3

  • eBook Packages: Springer Protocols

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