Doctor of Philosophy (PhD)
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TOF-SIMS, Mass Spectrometry Imaging, Subcellular Chemical Mapping, Imaging, MSI
Date of Defense
Time-of-Flight Secondary Ion Mass Spectrometry (TOF-SIMS) is a powerful mass spectrometry imaging technique that is uniquely suited to the study of small and challenging biological samples. Mass spectrometry imaging permits the simultaneous detection of chemical species without a priori knowledge of a sample. In the present work, the use of endogenous chemical species, three-dimensional dual-beam depth profiling, and multi-modal mass spectrometry approaches are utilized to increase the specificity and reliability of sub-cellular chemical assignments. In addition, the use of these complimentary indicators serve as useful quality controls to ensure optimal preparation of samples for a high-vacuum environment. In the study of gunshot residue, the simultaneous detection of ‘characteristic’ inorganic and organic components using TOF-SIMS combined with secondary electron imaging from a particulate enabled confirmation of gunshot origin in a single analysis. The use of high spatial resolution TOF-SIMS imaging (~250 nm) permitted, for the first time, the localization and quantification of anti-tumor drug inside glioblastoma cells at therapeutic levels. The study of mosquito ovarian follicles from Aedes aegypti as a function of diet using 3D-TOF-SIMS imaging permitted, for the first time, the identification of endogenous molecular components within the oocyte, nurse cells, and follicular cells. When complemented with stable isotope labeling, 3D-TOF-SIMS allowed for the first time the mapping of de novo synthesized triacylglycerides as a function diet during the previtellogenic resting stage in mosquitoes; parallel studies using LC-TIMS-MS/MS permitted the identification and quantification of de novo synthesized triacylglycerides as a function of diet and time after eclosion. Lastly, a multi-modal, in situ imaging protocol is described for the first time capable of protein/small molecule characterization and localization of biomarkers associated with human acute wound healing.
Previously Published In
- A. Castellanos; C.E. Ramirez; V. Michalkova; M. Nouzova; F.G. Noriega; F. Fernandez-Lima, Three dimensional secondary ion mass spectrometry imaging (3D-SIMS) of Aedes aegypti ovarian follicles, J. Anal. At. Spectrom., 2019. DOI: 10.1039/c8ja00425k.
- Q. Vanbellingen; A. Castellanos; M. Rodriguez Silva; I. Paudel, J. Chambers; F. Fernandez-Lima, Analysis of Chemotherapeutic Drug Delivery at the Single Cell level using 3D-MSI-TOF-SIMS, JASMS 27, 2033-2040, 2016. DOI: 10.1007/s13361-016-1485-y
- A. Castellanos; S. Bell; F. Fernandez-Lima, Characterization of Firearms Discharge Residue Recovered from Skin Swabs using Sub-micrometric Mass Spectrometry Imaging, Anal. Methods 8, 4300-4305, 2016. DOI: 10.1039/c6ay00096g
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 License.
Castellanos, Anthony, "Topological Markers for Subcellular Chemical Mapping using TOF-SIMS" (2019). FIU Electronic Theses and Dissertations. 4337.
Available for download on Friday, October 15, 2021
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