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Cryo-OrbiSIMS for 3D molecular imaging of a bacterial biofilm in its native state

Junting Zhang, James Brown, David Scurr, Anwen Bullen, Kirsty MacLellan-Gibson, Paul Williams, Morgan R. Alexander, Kim R. Hardie, Ian S. Gilmore, Paulina D. Rakowska
doi: https://doi.org/10.1101/859967
Junting Zhang
1National Physical Laboratory, NiCE-MSI, Teddington, Middlesex, UK
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James Brown
2Centre for Biomolecular Sciences and School of Life Sciences, The University of Nottingham, Nottingham, UK
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David Scurr
3School of Pharmacy, The University of Nottingham, Nottingham, UK
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Anwen Bullen
4National Institute for Biological Standards and Controls, Biological Imaging Group, Potters Bar, UK
5UCL Ear Institute, London, UK
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Kirsty MacLellan-Gibson
4National Institute for Biological Standards and Controls, Biological Imaging Group, Potters Bar, UK
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Paul Williams
2Centre for Biomolecular Sciences and School of Life Sciences, The University of Nottingham, Nottingham, UK
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Morgan R. Alexander
3School of Pharmacy, The University of Nottingham, Nottingham, UK
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Kim R. Hardie
2Centre for Biomolecular Sciences and School of Life Sciences, The University of Nottingham, Nottingham, UK
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Ian S. Gilmore
1National Physical Laboratory, NiCE-MSI, Teddington, Middlesex, UK
3School of Pharmacy, The University of Nottingham, Nottingham, UK
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  • For correspondence: ian.gilmore@npl.co.uk
Paulina D. Rakowska
1National Physical Laboratory, NiCE-MSI, Teddington, Middlesex, UK
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Abstract

We describe a method for label-free molecular imaging of biological materials, preserved in a native state, by using an OrbiSIMS instrument equipped with cryogenic sample handling and developing a high-pressure freezing protocol compatible with mass spectrometry. We studied the 3D distribution of quorum sensing signalling molecules, nucleobases and bacterial membrane molecules, in a mature Pseudomonas aeruginosa biofilm, with high spatial-resolution and high mass-resolution.

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  • ↵† Senior author at institute

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Posted November 30, 2019.
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Cryo-OrbiSIMS for 3D molecular imaging of a bacterial biofilm in its native state
Junting Zhang, James Brown, David Scurr, Anwen Bullen, Kirsty MacLellan-Gibson, Paul Williams, Morgan R. Alexander, Kim R. Hardie, Ian S. Gilmore, Paulina D. Rakowska
bioRxiv 859967; doi: https://doi.org/10.1101/859967
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Cryo-OrbiSIMS for 3D molecular imaging of a bacterial biofilm in its native state
Junting Zhang, James Brown, David Scurr, Anwen Bullen, Kirsty MacLellan-Gibson, Paul Williams, Morgan R. Alexander, Kim R. Hardie, Ian S. Gilmore, Paulina D. Rakowska
bioRxiv 859967; doi: https://doi.org/10.1101/859967

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