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Cell-Based Imaging Assay for Detection of Phospholipidosis

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High-Throughput Screening Assays in Toxicology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2474))

Abstract

Accumulation of lysosomal phospholipids in cells exposed to cationic amphiphilic drugs is characteristic of drug-induced phospholipidosis. The morphological hallmark of phospholipidosis is the appearance of unicentric or multicentric-lamellar bodies when viewed under an electron microscope (EM). The EM method, the gold standard of detecting cellular phospholipidosis, has downsides, namely, low-throughput, high-costs, and unsuitability for screening a large chemical library. This chapter describes a cell-based high-content phospholipidosis assay using the LipidTOX reagent in a high-throughput screening (HTS) platform. This assay has been optimized and validated in HepG2 and HepRG cells, and miniaturized into a 1536-well plate, thus can be used for high-throughput screening (HTS) to identify chemical compounds that induce phospholipidosis.

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Acknowledgments

This work was supported, in part, by the Intramural research program of the NCATS, NIH.

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Authors and Affiliations

  1. National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, USA

    Li Zhang, Shuaizhang Li & Menghang **a

Authors
  1. Li Zhang
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  2. Shuaizhang Li
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  3. Menghang **a
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Corresponding author

Correspondence to Menghang **a .

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Editors and Affiliations

  1. Center for Computational and Integrative Biology, Rutgers University, Camden, NJ, USA

    Hao Zhu

  2. National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, USA

    Menghang **a

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Zhang, L., Li, S., **a, M. (2022). Cell-Based Imaging Assay for Detection of Phospholipidosis. In: Zhu, H., **a, M. (eds) High-Throughput Screening Assays in Toxicology. Methods in Molecular Biology, vol 2474. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2213-1_8

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  • DOI: https://doi.org/10.1007/978-1-0716-2213-1_8

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2212-4

  • Online ISBN: 978-1-0716-2213-1

  • eBook Packages: Springer Protocols

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