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Single cell label-free probing of chromatin dynamics during B lymphocyte maturation

Rikke Morrish, Kevin Ho Wai Yim, Stefano Pagliara, Francesca Palombo, View ORCID ProfileRichard Chahwan, Nicholas Stone
doi: https://doi.org/10.1101/2021.01.12.426344
Rikke Morrish
1School of Physics and Astronomy, University of Exeter, Exeter, EX4 4QL, United Kingdom
2Living Systems Institute and School of Biosciences, University of Exeter, Exeter, United Kingdom
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Kevin Ho Wai Yim
3Institute of Experimental Immunology, University of Zurich, Zurich, Switzerland
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Stefano Pagliara
2Living Systems Institute and School of Biosciences, University of Exeter, Exeter, United Kingdom
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  • For correspondence: s.pagliara@exeter.ac.uk f.palombo@exeter.ac.uk chahwan@immunology.uzh.ch n.stone@exeter.ac.uk
Francesca Palombo
1School of Physics and Astronomy, University of Exeter, Exeter, EX4 4QL, United Kingdom
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  • For correspondence: s.pagliara@exeter.ac.uk f.palombo@exeter.ac.uk chahwan@immunology.uzh.ch n.stone@exeter.ac.uk
Richard Chahwan
3Institute of Experimental Immunology, University of Zurich, Zurich, Switzerland
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  • ORCID record for Richard Chahwan
  • For correspondence: s.pagliara@exeter.ac.uk f.palombo@exeter.ac.uk chahwan@immunology.uzh.ch n.stone@exeter.ac.uk
Nicholas Stone
1School of Physics and Astronomy, University of Exeter, Exeter, EX4 4QL, United Kingdom
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  • For correspondence: s.pagliara@exeter.ac.uk f.palombo@exeter.ac.uk chahwan@immunology.uzh.ch n.stone@exeter.ac.uk
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ABSTRACT

Large-scale intracellular signalling during developmental growth or in response to environmental alterations are largely orchestrated by chromatin within the cell nuclei. Chemical and conformational modifications of the chromatin architecture are critical steps in the regulation of differential gene expression and ultimately cell fate determination. Therefore, establishing chemical properties of the nucleus could provide key markers for phenotypic characterisation of cellular processes on a scale of individual cells.

Raman microscopy is a sensitive technique that is capable of probing single cell chemical composition - and sub-cellular regions - in a label-free optical manner. As such, it has great potential in both clinical and basic research. However, perceived limitations of Raman spectroscopy such as low signal intensity and the difficulty in linking alterations in vibrational signals directly with ensuing biological effects have hampered advances in the field. Here we use immune B lymphocyte development as a model to assess chromatin and transcriptional changes using confocal Raman microscopy in combination with microfluidic devices and correlative transcriptomics, thereby linking changes in chemical and structural properties to biological outcomes. Live B lymphocytes were assessed before and after maturation. Multivariate analysis was applied to distinguish cellular components within each cell. The spectral differences between non-activated and activated B lymphocytes were then identified, and their correlation with known intracellular biological changes were assessed in comparison to conventional RNA-seq analysis. Our data shows that spectral analysis provides a powerful tool to study gene activation that can complement conventional molecular biology techniques and opens the way for mapping the dynamics in the biochemical makeup of individual cells.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted January 13, 2021.
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Single cell label-free probing of chromatin dynamics during B lymphocyte maturation
Rikke Morrish, Kevin Ho Wai Yim, Stefano Pagliara, Francesca Palombo, Richard Chahwan, Nicholas Stone
bioRxiv 2021.01.12.426344; doi: https://doi.org/10.1101/2021.01.12.426344
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Single cell label-free probing of chromatin dynamics during B lymphocyte maturation
Rikke Morrish, Kevin Ho Wai Yim, Stefano Pagliara, Francesca Palombo, Richard Chahwan, Nicholas Stone
bioRxiv 2021.01.12.426344; doi: https://doi.org/10.1101/2021.01.12.426344

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