Zhang, J;
Brown, J;
Scurr, DJ;
Bullen, A;
MacLellan-Gibson, K;
Williams, P;
Alexander, MR;
... Rakowska, PD; + view all
(2020)
Cryo-OrbiSIMS for 3D Molecular Imaging of a Bacterial Biofilm in Its Native State.
Analytical Chemistry
, 92
(13)
pp. 9008-9015.
10.1021/acs.analchem.0c01125.
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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.
| Type: | Article |
|---|---|
| Title: | Cryo-OrbiSIMS for 3D Molecular Imaging of a Bacterial Biofilm in Its Native State |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1021/acs.analchem.0c01125 |
| Publisher version: | https://doi.org/10.1021/acs.analchem.0c01125 |
| Language: | English |
| Additional information: | This version is the author accepted manuscript. For information on re-use, please refer to the publisher's terms and conditions. |
| Keywords: | Biofilms, Lipids, Mass spectrometry, Molecules, Ions |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > The Ear Institute |
| URI: | https://discovery-pp.ucl.ac.uk/id/eprint/10108018 |
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