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

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

Secondary ion mass spectrometry (SIMS) is gaining popularity for molecular imaging in the life sciences because it is label-free and allows imaging in two and three dimensions. The recent introduction of the OrbiSIMS has significantly improved the utility for biological imaging through combining subcellular spatial resolution with high-performance Orbitrap mass spectrometry. SIMS instruments operate in high-vacuum, and samples are typically analyzed in a freeze-dried state. Consequently, the molecular and structural information may not be well-preserved. We report a method for molecular imaging of biological materials, preserved in a native state, by using an OrbiSIMS instrument equipped with cryogenic sample handling and a high-pressure freezing protocol compatible with mass spectrometry. The performance is demonstrated by imaging a challenging sample (>90% water) of a mature Pseudomonas aeruginosa biofilm in its native state. The 3D distribution of quorum sensing signaling molecules, nucleobases, and bacterial membrane molecules is revealed with high spatial-resolution and high mass-resolution. We discover that analysis in the frozen-hydrated state yields a 10 »000-fold increase in signal intensity for polar molecules such as amino acids, which has important implications for SIMS imaging of metabolites and pharmaceuticals.

0003-2700
9008-9015
Zhang, Junting
86ce6187-513a-4d8a-81be-59f8228696f8
Brown, James
9d3abf07-c43e-4a01-a3bc-4dfe26b26101
Scurr, David J.
9cf743e6-6247-4116-8732-7e2cd8d934ed
Bullen, Anwen
97f74994-b4c1-4e47-837d-66528f4ff4d8
MacLellan-Gibson, Kirsty
90c04653-6a4e-4f53-aebb-4ee6906f2081
Williams, Paul
fadf7f35-ba19-405d-9346-e976004c589f
Alexander, Morgan R.
d252e635-239e-4e68-8a07-eaf7bac0eb93
Hardie, Kim R.
9a63b899-ba6d-422f-8e1f-93193a125798
Gilmore, Ian S.
f864335b-9cb0-4f44-b80f-30809227212b
Rakowska, Paulina D.
73bf2145-e2fa-46ee-9ad9-f74c80d5a369
Zhang, Junting
86ce6187-513a-4d8a-81be-59f8228696f8
Brown, James
9d3abf07-c43e-4a01-a3bc-4dfe26b26101
Scurr, David J.
9cf743e6-6247-4116-8732-7e2cd8d934ed
Bullen, Anwen
97f74994-b4c1-4e47-837d-66528f4ff4d8
MacLellan-Gibson, Kirsty
90c04653-6a4e-4f53-aebb-4ee6906f2081
Williams, Paul
fadf7f35-ba19-405d-9346-e976004c589f
Alexander, Morgan R.
d252e635-239e-4e68-8a07-eaf7bac0eb93
Hardie, Kim R.
9a63b899-ba6d-422f-8e1f-93193a125798
Gilmore, Ian S.
f864335b-9cb0-4f44-b80f-30809227212b
Rakowska, Paulina D.
73bf2145-e2fa-46ee-9ad9-f74c80d5a369

Zhang, Junting, Brown, James, Scurr, David J., Bullen, Anwen, MacLellan-Gibson, Kirsty, Williams, Paul, Alexander, Morgan R., Hardie, Kim R., Gilmore, Ian S. and Rakowska, Paulina D. (2020) Cryo-OrbiSIMS for 3D molecular imaging of a bacterial biofilm in its native state. Analytical Chemistry, 92 (13), 9008-9015. (doi:10.1021/acs.analchem.0c01125).

Record type: Article

Abstract

Secondary ion mass spectrometry (SIMS) is gaining popularity for molecular imaging in the life sciences because it is label-free and allows imaging in two and three dimensions. The recent introduction of the OrbiSIMS has significantly improved the utility for biological imaging through combining subcellular spatial resolution with high-performance Orbitrap mass spectrometry. SIMS instruments operate in high-vacuum, and samples are typically analyzed in a freeze-dried state. Consequently, the molecular and structural information may not be well-preserved. We report a method for molecular imaging of biological materials, preserved in a native state, by using an OrbiSIMS instrument equipped with cryogenic sample handling and a high-pressure freezing protocol compatible with mass spectrometry. The performance is demonstrated by imaging a challenging sample (>90% water) of a mature Pseudomonas aeruginosa biofilm in its native state. The 3D distribution of quorum sensing signaling molecules, nucleobases, and bacterial membrane molecules is revealed with high spatial-resolution and high mass-resolution. We discover that analysis in the frozen-hydrated state yields a 10 »000-fold increase in signal intensity for polar molecules such as amino acids, which has important implications for SIMS imaging of metabolites and pharmaceuticals.

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More information

Accepted/In Press date: 28 May 2020
e-pub ahead of print date: 28 May 2020
Published date: 7 July 2020

Identifiers

Local EPrints ID: 445197
URI: http://eprints.soton.ac.uk/id/eprint/445197
ISSN: 0003-2700
PURE UUID: 7d859fb4-821d-48c6-a89d-f562ed570910
ORCID for Paulina D. Rakowska: ORCID iD orcid.org/0000-0002-3710-8395

Catalogue record

Date deposited: 25 Nov 2020 17:30
Last modified: 28 Apr 2022 02:31

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Contributors

Author: Junting Zhang
Author: James Brown
Author: David J. Scurr
Author: Anwen Bullen
Author: Kirsty MacLellan-Gibson
Author: Paul Williams
Author: Morgan R. Alexander
Author: Kim R. Hardie
Author: Ian S. Gilmore

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