Exploring Chromosome Architecture Using Multi-modal Imaging

Session

Session 1 - Multimodal Imaging including Correlative

Authors

Daniel Booth (2), Eva Simpson (2), Alison Beckett (1), Ian Prior (1)

Affiliations

1. University of Liverpool
2. University of Nottingham

Keywords

CLEM, 3DCLEM, chromosomes, mitosis.

Abstract text

Since their first discovery in 1882, chromosomes have been a subject of intense study. Remarkably, despite the significant developments of light (LM) and electron microscopy (EM) over the intervening years, the detailed organization of condensed chromosomes and in particular how this might change throughout the stages of cell division, remains a mystery. Defining the fundamental properties of chromosomes is important, as defects in chromosome structure, organisation and composition can lead to aneuploidy, a feature of many cancers, birth defects, and linked to infertility and miscarriage. 

To address this gap in understanding, we have established a robust 3D correlative light electron microscopy (3D-CLEM) pipeline to assemble the first “chromosome atlas” – an ultrastructural survey of all chromosomes, during each stage of cell division. This atlas can now act as a reference map for “normal” cell division and used to study and compare samples/datasets with suspected chromosome anomalies. 

We also show how the same tools can be adapted to investigate a “needle in the haystack” – that is, small areas of interest found within large and precious cells. 

We hope these advances are not only useful for the chromosome biology community but also the wider cell biology field. 

References

Booth, D. G^., Beckett, A. J., Meijer D. M^. 2019. SuperCLEM: An accessible correlative light and electron microscopy approach for investigation of neurons and glia in vitro. Biology Open.

Samejima, K*., Booth, D. G*., Paulson, J. R., Earnshaw, W.C. 2018. Functional analysis after rapid degradation of condensins and 3D-EM reveals chromatin volume is uncoupled from chromosome architecture in mitosis. J Cell Sci, 131, 210181.

Booth, D. G^., Beckett, A. J., Molina, O., Masumoto, H., Kouprina, K., Larionov, V., Prior, I. A., Earnshaw, W. C^. 2016. 3D-CLEM Reveals that a Major Portion of Mitotic Chromosomes Is Not Chromatin. Molecular Cell, 64,1-13.