Cryo-OrbiSIMS for 3D molecular imaging of a bacterial biofilm in its native state
Creators
- 1. National Physical Laboratory, NiCE-MSI
- 2. Biodiscovery Institute and School of Life Sciences, The University of Nottingham
- 3. School of Pharmacy, The University of Nottingham
- 4. UCL Ear Institute
- 5. National Institute for Biological Standards and Controls, Biological Imaging Group
Description
We developed a method for analysis and imaging of biological samples in their native state, by combining a cryo-OrbiSIMS instrument with cryogenic sample handling and high-pressure freezing. By using this method, we did analysis and imaging of frozen-hydrated mature Pseudomonas aeruginosa biofilm, which allows the identification and map of quorum sensing signaling molecules, nucleobases and bacterial membrane molecules with high spatial-resolution and high mass-resolution. Some of quorum sensing signaling molecules were further confirmed by MS/MS. By comparing the analysis of frozen-hydrated Pseudomonas aeruginosa biofilm with the freeze-dried one, we dicover that signal intensity of all interesting molecules get enhanced in the frozen-hydrated state. Especially for polar molecules, such as amino acid, it could even achieve 10,000 fold increasing. Here, we provide the original OrbiSIMS data including MS and MS/MS spectra, depth profile and images of frozen-hydrated and freeze-dried Pseudomonas aeruginosa biofilm. The data could be open by using SurfaceLab Version 7.0 (ION-TOF, Germany).
Notes
Files
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Additional details
Funding
- UK Research and Innovation
- 3D OrbiSIMS: Label free chemical imaging of materials, cells and tissues EP/P029868/1