Measuring Cellular Ploidy In Situ by Light Microscopy.

Autor:	Clay DE; Department of Pharmacology and Cancer Biology, Duke University Medical Center, Durham, NC, USA., Stormo BM; Department of Pharmacology and Cancer Biology, Duke University Medical Center, Durham, NC, USA., Fox DT; Department of Pharmacology and Cancer Biology, Duke University Medical Center, Durham, NC, USA. don.fox@duke.edu.
Jazyk:	angličtina
Zdroj:	Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2023; Vol. 2545, pp. 401-412.
DOI:	10.1007/978-1-0716-2561-3_21
Abstrakt:	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. (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu