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Measuring Cellular Ploidy In Situ by Light Microscopy.

Publication ,  Journal Article
Clay, DE; Stormo, BM; Fox, DT
Published in: Methods Mol Biol
2023

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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Published In

Methods Mol Biol

DOI

EISSN

1940-6029

Publication Date

2023

Volume

2545

Start / End Page

401 / 412

Location

United States

Related Subject Headings

  • Ploidies
  • Microscopy
  • Drosophila melanogaster
  • Drosophila
  • Developmental Biology
  • DNA
  • Cell Nucleus
  • Animals
  • 3404 Medicinal and biomolecular chemistry
  • 3101 Biochemistry and cell biology
 

Citation

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Clay, D. E., Stormo, B. M., & Fox, D. T. (2023). Measuring Cellular Ploidy In Situ by Light Microscopy. Methods Mol Biol, 2545, 401–412. https://doi.org/10.1007/978-1-0716-2561-3_21
Clay, Delisa E., Benjamin M. Stormo, and Donald T. Fox. “Measuring Cellular Ploidy In Situ by Light Microscopy.Methods Mol Biol 2545 (2023): 401–12. https://doi.org/10.1007/978-1-0716-2561-3_21.
Clay DE, Stormo BM, Fox DT. Measuring Cellular Ploidy In Situ by Light Microscopy. Methods Mol Biol. 2023;2545:401–12.
Clay, Delisa E., et al. “Measuring Cellular Ploidy In Situ by Light Microscopy.Methods Mol Biol, vol. 2545, 2023, pp. 401–12. Pubmed, doi:10.1007/978-1-0716-2561-3_21.
Clay DE, Stormo BM, Fox DT. Measuring Cellular Ploidy In Situ by Light Microscopy. Methods Mol Biol. 2023;2545:401–412.

Published In

Methods Mol Biol

DOI

EISSN

1940-6029

Publication Date

2023

Volume

2545

Start / End Page

401 / 412

Location

United States

Related Subject Headings

  • Ploidies
  • Microscopy
  • Drosophila melanogaster
  • Drosophila
  • Developmental Biology
  • DNA
  • Cell Nucleus
  • Animals
  • 3404 Medicinal and biomolecular chemistry
  • 3101 Biochemistry and cell biology