A novel tool for neuronal physiology and pathology: optimization of secondary ion mass spectrometry approaches

von Paola Agüí González
Dissertation
Datum der mündl. Prüfung:2023-05-11
Erschienen:2023-10-05
Betreuer:Prof. Dr. Silvio O. Rizzoli
Gutachter:Prof. Dr. Silvio O. Rizzoli
Gutachter:Prof. Dr. Peter Rehling
Gutachter:Prof. Dr. Thomas Meyer
Zum Verlinken/Zitieren: http://dx.doi.org/10.53846/goediss-9903

 

 

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Description:Doctoral Thesis - Paola Agüí González

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Zusammenfassung

Englisch

Secondary Ion Mass Spectrometry (SIMS) is a powerful tool for characterizing the chemical composition of solid samples and thin films, offering high sensitivity and spatial resolution. While initially developed for geology, cosmochemistry and material science, the continuous evolution of this imaging technique has broadened its scope towards other disciplines, including biology. Despite inherent challenges and limitations in SIMS imaging, through the four projects included in this thesis, I demonstrate that it is possible to perform quantitative biological imaging with this technique, overcoming some of the most important limitations by introducing novel tools and combining SIMS with other imaging techniques. First, Time of Flight Mass Secondary Ion Mass Spectrometry (ToF-SIMS) was employed to investigate the potential correlation between the synaptic activity of hippocampal neurons and the composition and distribution of lipid species on their plasma membranes. Second, I present some novel probes, detectable with both SIMS and fluorescence microscopy, which enabled protein localization at the subcellular level. Third, Nanoscale Secondary Ion Mass Spectrometry (NanoSIMS) was correlated with Transmission Electron Microscopy (TEM), to investigate the inclusion of newly synthesized proteins into mature myelin sheaths. Fourth, we developed and successfully applied a protocol to correlate three imaging techniques. Through a single experiment, this protocol enables the localization of specific targets with light microscopy, the analysis of the morphology of cellular structures with TEM, and the investigation of the chemical composition of regions of interest with NanoSIMS. In summary, the work included in this thesis aims to boost the use of SIMS imaging in neuro- and cell biology, showing how the combination of this technique with new tools and technologies has the potential to enhance our knowledge about cellular mechanisms, as well as facilitating diagnostics and drug development.
Keywords: SIMS imaging; Image correlation; Cell biology; Analytical technique
Schlagwörter: SIMS; Image correlation; Analytical technique; Cell biology
 
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