title: ToF-SIMS of tissues: "lessons learned" from mice and women.
abstract: The ability to image cells and tissues with chemical and molecular specificity could greatly expand our understanding of biological processes. The subcellular resolution mass spectral imaging capability of time of flight secondary ion mass spectrometry (ToF-SIMS) has the potential to acquire chemically detailed images. However, the complexities of biological systems combined with the sensitivity of ToF-SIMS require careful planning of experimental methods. Tissue sample preparation methods of formalin fixation followed by paraffin embedding (FFPE) and OCT embedding are compared. Results show that the FFPE can potentially be used as a tissue sample preparation protocol for ToF-SIMS analysis if a cluster ion pre-sputter is used prior to analysis and if nonlipid related tissue features are the features of interest. In contrast, embedding tissue in OCT minimizes contamination and maintains lipid signals. Various data acquisition methodologies and analysis options are discussed and compared using mouse breast and diaphragm muscle tissue. Methodologies for acquiring ToF-SIMS 2D images are highlighted along with applications of multivariate analysis to better identify specific features in a tissue sections when compared to H&E images of serial sections. Identification of tissue features is necessary for researchers to visualize a molecular map that correlates with specific biological features or functions. Finally, lessons learned from sample preparation, data acquisition, and data analysis methods developed using mouse models are applied to a preliminary analysis of human breast tumor tissue sections.
authors.affiliation: Department of Bioengineering, Molecular Engineering and Sciences Building, University of Washington, Box 351653, Seattle, Washington 98195-1653.
authors.forename: Lara J
authors.initials: LJ
authors.lastname: Gamble
authors.affiliation: Department of Bioengineering, Molecular Engineering and Sciences Building, University of Washington, Box 351653, Seattle, Washington 98195-1653.
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Graham
authors.affiliation: Department of Bioengineering, Molecular Engineering and Sciences Building, University of Washington, Box 351653, Seattle, Washington 98195-1653.
authors.forename: Blake
authors.initials: B
authors.lastname: Bluestein
authors.affiliation: Department of Physiology and Biophysics, University of Washington, Box 357290, Seattle, Washington 98195-1653.
authors.forename: Nicholas P
authors.initials: NP
authors.lastname: Whitehead
authors.affiliation: Fred Hutchinson Cancer Research Center, Seattle, Washington 98109.
authors.forename: David
authors.initials: D
authors.lastname: Hockenbery
authors.affiliation: Fred Hutchinson Cancer Research Center, Seattle, Washington 98109.
authors.forename: Fionnuala
authors.initials: F
authors.lastname: Morrish
authors.affiliation: Fred Hutchinson Cancer Research Center, Seattle, Washington 98109.
authors.forename: Peggy
authors.initials: P
authors.lastname: Porter
mesh_terms: D000818:Animals; D001943:Breast Neoplasms / Q000473:pathology; D003964:Diaphragm / Q000473:pathology; D005260:Female; D006801:Humans; D008321:Mammary Glands, Animal / Q000473:pathology; D051379:Mice; D010336:Pathology / Q000379:methods*; D018629:Spectrometry, Mass, Secondary Ion / Q000379:methods*; D016610:Tissue Embedding / Q000379:methods*; D016707:Tissue Fixation / Q000379:methods*