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DEVELOPMENTS IN AMBIENT MASS SPECTROMETRY IMAGING FOR IN-DEPTH SPATIALLY RESOLVED ANALYSIS OF COMPLEX BIOLOGICAL TISSUES
Ambient Mass Spectrometry Imaging (MSI) is a powerful analytical tool in biomedical research that enables simultaneous label-free spatial mapping of hundreds of molecules in biological samples under native conditions. Nanospray desorption electrospray ionization (nano-DESI) is an emergent ambient MSI technique developed in 2010 that uses localized liquid extraction of molecules directly from surfaces. Like other liquid-extraction based techniques, nano-DESI relies on gentle removal of molecules from surfaces and soft ionization. High sensitivity and spatial resolution, versatility of the solvent composition, which may be used to tailor the extraction and ionization of selected molecules, quantification capabilities at the single-pixel level as well as compensation for matrix effects by adding a known standard to the solvent, and online derivatization are key features of nano-DESI MSI that position it as a unique analytical tool for studying biological systems.
Despite the advantages that nano-DESI provides, there are still challenges associated with the structural characterization, extraction, and detection of certain molecular classes. Therefore, my dissertation research has focused on addressing these analytical challenges by developing innovative approaches that substantially enhance the performance of the nano-DESI technique in the study of complex biological systems.
In this thesis, a systematic study of the solvent composition is carried out to aid in the detection of neutral lipids such as triglycerides thereby expanding the molecular coverage of nano-DESI experiments. Taking advantage of the versatility of the solvent composition, I developed an approach for the online derivatization of unsaturated lipids into lipid hydroperoxides using the reaction of singlet oxygen with C=C bonds. This method further expands the specificity of nano-DESI MSI by enabling the detection and imaging of positional lipid isomers. To aid in the analysis of complex mixtures and provide additional structural information in the form of collision cross sections, coupling of nano-DESI with a drift-tube ion mobility spectrometry is also reported along with examples of the powerful capabilities of this platform. Lastly, nano-DESI MSI is used to address the complexity in the analysis of individual skeletal muscle fibers. This collaborative project involves the development of a robust image registration approach of immunofluorescence imaging and high-spatial resolution nano-DESI MSI to obtain accurate chemical maps specific to each fiber type. The developments described in this thesis are key to understanding the dynamic metabolic processes on a molecular level with an unprecedented specificity and sensitivity.
- Doctor of Philosophy
- West Lafayette