Segmentation, 3D Reconstruction, and Analysis of PcG Proteins in Fluorescence Microscopy Images in Different Cell Culture Conditions

Methods Mol Biol. 2023:2655:147-169. doi: 10.1007/978-1-0716-3143-0_12.

Abstract

Polycomb-group (PcG) of proteins are evolutionarily conserved transcription factors necessary for the regulation of gene expression during the development and the safeguard of cell identity in adulthood. In the nucleus, they form aggregates whose positioning and dimension are fundamental for their function. We present an algorithm, and its MATLAB implementation, based on mathematical methods to detect and analyze PcG proteins in fluorescence cell image z-stacks. Our algorithm provides a method to measure the number, the size, and the relative positioning of the PcG bodies in the nucleus for a better understanding of their spatial distribution, and thus of their role for a correct genome conformation and function.

Keywords: Cellular and subcellular segmentation; Fluorescence microscopy; Image processing and analysis; Nuclear organization; PcG staining; Unsupervised classification algorithm; Variational segmentation model.

MeSH terms

  • Cell Culture Techniques
  • Cell Nucleus* / metabolism
  • Imaging, Three-Dimensional*
  • Microscopy, Fluorescence / methods
  • Polycomb-Group Proteins

Substances

  • Polycomb-Group Proteins