Quantitative imaging of chromatin inheritance using a dual-color histone in Drosophila germinal stem cells

STAR Protoc. 2022 Nov 2;3(4):101811. doi: 10.1016/j.xpro.2022.101811. eCollection 2022 Dec 16.

Abstract

We detail a stepwise protocol for the investigation and quantification of chromatin features during asymmetric cell division (ACD) of Drosophila germline stem cells (GSCs). We describe the use of a dual-color histone to study the inheritance of new and old histones. We detail steps for imaging and analysis of sister chromatid condensation dynamics and nucleosome density changes. In addition, this protocol could be applied to identify stem cells, which can be challenging to identify in intact tissues. For complete details on the use and execution of this protocol, please refer to Tran et al. (2012), Ranjan et al. (2019), and Ranjan et al. (2022).

Keywords: Cell biology; Developmental biology; Microscopy; Model organisms; Molecular biology; Molecular/Chemical probes; Stem cells.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chromatin / genetics
  • Drosophila Proteins* / genetics
  • Drosophila melanogaster
  • Drosophila* / genetics
  • Germ Cells
  • Histones / genetics
  • Stem Cells

Substances

  • Histones
  • Chromatin
  • Drosophila Proteins