期刊
ANALYTICAL CHEMISTRY
卷 92, 期 13, 页码 9008-9015出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.0c01125
关键词
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资金
- MetVBadBugs project of the EMPIR programme [15HLTH01]
- European Union's Horizon 2020 research and innovation programme
- EPSRC [EP/P029868/1]
- BBSRC [BB/R012415/1] Funding Source: UKRI
- EPSRC [EP/P029868/1] Funding Source: UKRI
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 spatialresolution 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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