Measuring nanoscale chromatin heterogeneity with partial wave spectroscopic microscopy

Scott Gladstein, Andrew Stawarz, Luay M. Almassalha, Lusik Cherkezyan, John E. Chandler, Xiang Zhou, Hariharan Subramanian, Vadim Backman*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Scopus citations

Abstract

Despite extensive research in the area, current understanding of the structural organization of higher-order chromatin topology (between 20 and 200 nm) is limited due to a lack of proper imaging techniques at these length scales. The organization of chromatin at these scales defines the physical context (nanoenvironment) in which many important biological processes occur. Improving our understanding of the nanoenvironment is crucial because it has been shown to play a critical functional role in the regulation of chemical reactions. Recent progress in partial wave spectroscopic (PWS) microscopy enables real-time measurement of higher-order chromatin organization within label-free live cells. Specifically, PWS quantifies the nanoscale variations in mass density (heterogeneity) within the cell. These advancements have made it possible to study the functional role of chromatin topology, such as its regulation of the global transcriptional state of the cell and its role in the development of cancer. In this chapter, the importance of studying chromatin topology is explained, the theory and instrumentation of PWS are described, the measurements and analysis processes for PWS are laid out in detail, and common issues, troubleshooting steps, and validation techniques are provided.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages337-360
Number of pages24
DOIs
StatePublished - Jan 1 2018

Publication series

NameMethods in Molecular Biology
Volume1745
ISSN (Print)1064-3745

Keywords

  • Cancer development
  • Chromatin structure
  • Chromatin topology
  • Fractal dimension
  • Heterogeneity
  • Live cell imaging
  • Nanoscale imaging
  • Partial wave spectroscopic microscopy
  • Transcriptional

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Gladstein, S., Stawarz, A., Almassalha, L. M., Cherkezyan, L., Chandler, J. E., Zhou, X., Subramanian, H., & Backman, V. (2018). Measuring nanoscale chromatin heterogeneity with partial wave spectroscopic microscopy. In Methods in Molecular Biology (pp. 337-360). (Methods in Molecular Biology; Vol. 1745). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-7680-5_19