Abstract
Determining cellular DNA content is valuable in the study of numerous biological processes, including organ development and injury repair. While FACS analysis of dissociated cells is a widely used method for assaying ploidy in a tissue cell population, for many tissue samples, it is possible and convenient to measure ploidy in situ using light microscopy. Here, we present two protocols for measuring cellular ploidy in tissues. These protocols are based on our studies in Drosophila melanogaster, but these are applicable to other settings as well. We present example results from Drosophila hindgut, midgut, and wing imaginal disc as examples. The first protocol focuses on measuring DNA content from decondensed interphase nuclei, while the second protocol details the visualization of condensed chromosomes for ploidy determination, either from mitotic cells or from interphase cells with drug-induced chromosome condensation. These techniques can be completed in 1 day and require standard lab supplies as well as a fluorescence light microscope.
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Acknowledgments
Study of polyploidy in the Fox laboratory is supported by a National Institutes of Health (NIH) grant R01GM118447. Delisa Clay is supported by a GRFP fellowship from the National Science Foundation (NSF). We thank Archan Chakraborty for comments on the manuscript.
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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
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Clay, D.E., Stormo, B.M., Fox, D.T. (2023). Measuring Cellular Ploidy In Situ by Light Microscopy. In: Van de Peer, Y. (eds) Polyploidy. Methods in Molecular Biology, vol 2545. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2561-3_21
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DOI: https://doi.org/10.1007/978-1-0716-2561-3_21
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