University of Illinois Urbana-Champaign

Applying secondary ion mass spectrometry to cellular and model membranes to image component distribution and quantify composition

Wilson, Robert

Content Files
Robert_Wilson.pdf

Permalink

https://hdl.handle.net/2142/42139

Description

Title
Applying secondary ion mass spectrometry to cellular and model membranes to image component distribution and quantify composition
Author(s)
Wilson, Robert
Issue Date
2013-02-03T19:16:57Z
Director of Research (if dissertation) or Advisor (if thesis)
Kraft, Mary L.
Doctoral Committee Chair(s)
Kraft, Mary L.
Committee Member(s)
  • Bailey, Ryan C.
  • Murphy, Catherine J.
  • Suslick, Kenneth S.
Department of Study
Chemistry
Discipline
Chemistry
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Date of Ingest
2013-02-03T19:16:57Z
Keyword(s)
  • Time-of-flight Secondary ion mass spectrometry (TOF-SIMS)
  • Lipid bilayers
  • NanoSIMS
  • Nanoparticle
  • Surface analysis
  • Multivariate analysis
  • Partial least squares regression
  • Spatial statistics
  • Stem cells
Abstract
Abstract Secondary ion mass spectrometry (SIMS) returns chemically specific data on membrane component distribution and composition. Of particular interest is a membrane protein’s lipid environment, which may play a role in the protein’s function. Herein, this thesis describes the synthesis of fluorine-functionalized colloidal gold immunolabels that facilitate the imaging of specific proteins in parallel with stable isotope labeled lipids in the cellular membrane using high-resolution SIMS performed with a NanoSIMS. Further, the spatial statistical analysis tools are outlined and applied to NanoSIMS data to test the hypothesis of colocalization of the flu virus protein hemagglutinin and sphingolipids. While the NanoSIMS has very high lateral resolution, it requires isotope labels for component identification. Time-of-flight SIMS (TOF-SIMS) detects large portions of the mass spectrum, allowing species to be identified by their fragmentation pattern. Ionization yields in TOF-SIMS, however, limit the lateral resolution and the maximum m/z that can be used consistently for analysis. These challenges can be addressed using multivariate analysis (MVA). TOF-SIMS and the MVA method partial least squares discriminant analysis (PLS-DA) was used to identify the differentiation stage of hematopoietic cells. Since TOF-SIMS is a location-specific technique, it can be used to investigate how varying chemical and material environments on one sample affect stem cell fate decisions. Another MVA method, partial least squares regression (PLSR), uses a calibrated model to return quantitative information from TOF-SIMS data. This was used to determine the concentration of cholesterol in homogeneous supported lipid membranes.
Graduation Semester
2012-12
Permalink
http://hdl.handle.net/2142/42139
Copyright and License Information
Copyright 2012 Robert Wilson

Owning Collections

Graduate Dissertations and Theses at Illinois PRIMARY
Graduate Theses and Dissertations at Illinois
Dissertations and Theses - Chemistry