Abstract
Fluorescent dyes have been used as “nanosensors” for visualization and determination of various processes occurring inside a cell, or intracellular events, such as cell cycle progression and intracellular trafficking. Here, we describe a novel use of acridine orange to visualize lysosomes and discriminate cells with healthy lysosomes from cells with damaged lysosomes in two different types of mammalian cells: fibroblasts and macrophages. This method allows assessment of lysosomal membrane integrity upon exposure to various foreign particles, i.e., engineered nanoparticles. The uniqueness of this method enables investigators to acquire fluorescent images with a dye that is susceptible to photo-bleaching under UV light. These acquired images bolster the quantitative data, providing a visual representation of the cell morphology as well as assess its nucleus and lysosomes.
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Acknowledgment
This work was supported by NIH grant EB007271.
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Sohaebuddin, S.K., Tang, L. (2013). A Simple Method to Visualize and Assess the Integrity of Lysosomal Membrane in Mammalian Cells Using a Fluorescent Dye. In: Weissig, V., Elbayoumi, T., Olsen, M. (eds) Cellular and Subcellular Nanotechnology. Methods in Molecular Biology, vol 991. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-336-7_3
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DOI: https://doi.org/10.1007/978-1-62703-336-7_3
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-335-0
Online ISBN: 978-1-62703-336-7
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