Measuring Cellular Ploidy In Situ by Light Microscopy.
Delisa E ClayBenjamin M StormoDonald T FoxPublished in: Methods in molecular biology (Clifton, N.J.) (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.
Keyphrases
- induced apoptosis
- single molecule
- drug induced
- cell cycle arrest
- liver injury
- randomized controlled trial
- high resolution
- drosophila melanogaster
- endoplasmic reticulum stress
- gene expression
- high speed
- signaling pathway
- circulating tumor
- oxidative stress
- cell proliferation
- cell free
- zika virus
- cell cycle
- single cell
- genome wide
- mass spectrometry
- mesenchymal stem cells
- bone marrow
- adverse drug