Document Type

Dissertation

Degree

Doctor of Philosophy (PhD)

Major/Program

Chemistry

First Advisor's Name

Francisco Fernandez-Lima

First Advisor's Committee Title

Committee Chair

Second Advisor's Name

Piero Gardinali

Second Advisor's Committee Title

Committee Member

Third Advisor's Name

Konstantinos Kavallieratos

Third Advisor's Committee Title

Committee Member

Fourth Advisor's Name

Jaroslava Miksovska

Fourth Advisor's Committee Title

Committee Member

Fifth Advisor's Name

Fernando Noriega

Fifth Advisor's Committee Title

Committee Member

Keywords

TOF-SIMS, Mass Spectrometry Imaging, Subcellular Chemical Mapping, Imaging, MSI

Date of Defense

11-6-2019

Abstract

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.

Identifier

FIDC008825

ORCID

0000-0003-3939-1507

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

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Available for download on Friday, October 15, 2021

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