title: Next-generation technologies for spatial proteomics: Integrating ultra-high speed MALDI-TOF and high mass resolution MALDI FTICR imaging mass spectrometry for protein analysis.
abstract: MALDI imaging mass spectrometry is a powerful analytical tool enabling the visualization of biomolecules in tissue. However, there are unique challenges associated with protein imaging experiments including the need for higher spatial resolution capabilities, improved image acquisition rates, and better molecular specificity. Here we demonstrate the capabilities of ultra-high speed MALDI-TOF and high mass resolution MALDI FTICR IMS platforms as they relate to these challenges. High spatial resolution MALDI-TOF protein images of rat brain tissue and cystic fibrosis lung tissue were acquired at image acquisition rates >25 pixels/s. Structures as small as 50 μm were spatially resolved and proteins associated with host immune response were observed in cystic fibrosis lung tissue. Ultra-high speed MALDI-TOF enables unique applications including megapixel molecular imaging as demonstrated for lipid analysis of cystic fibrosis lung tissue. Additionally, imaging experiments using MALDI FTICR IMS were shown to produce data with high mass accuracy (<5 ppm) and resolving power (∼75 000 at m/z 5000) for proteins up to ∼20 kDa. Analysis of clear cell renal cell carcinoma using MALDI FTICR IMS identified specific proteins localized to healthy tissue regions, within the tumor, and also in areas of increased vascularization around the tumor.
authors.affiliation: Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN, USA.|Mass Spectrometry Research Center, Vanderbilt University School of Medicine, Nashville, TN, USA.|Department of Chemistry, Vanderbilt University, Nashville, TN, USA.
authors.forename: Jeffrey M
authors.initials: JM
authors.lastname: Spraggins
authors.affiliation: Mass Spectrometry Research Center, Vanderbilt University School of Medicine, Nashville, TN, USA.|Department of Chemistry, Vanderbilt University, Nashville, TN, USA.
authors.forename: David G
authors.initials: DG
authors.lastname: Rizzo
authors.affiliation: Mass Spectrometry Research Center, Vanderbilt University School of Medicine, Nashville, TN, USA.|Department of Chemistry, Vanderbilt University, Nashville, TN, USA.
authors.forename: Jessica L
authors.initials: JL
authors.lastname: Moore
authors.affiliation: Department of Medicine, Vanderbilt University Medical Center, Nashville, TN, USA.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Noto
authors.affiliation: Department of Pathology, Microbiology and Immunology, Vanderbilt University School of Medicine, Nashville, TN, USA.|United States (U.S.) Department of Veterans Affairs, Tennessee Valley Healthcare System, Nashville, TN, USA.
authors.forename: Eric P
authors.initials: EP
authors.lastname: Skaar
authors.affiliation: Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN, USA.|Mass Spectrometry Research Center, Vanderbilt University School of Medicine, Nashville, TN, USA.|Department of Chemistry, Vanderbilt University, Nashville, TN, USA.|Department of Medicine, Vanderbilt University Medical Center, Nashville, TN, USA.|Departments of Pharmacology and Medicine, Vanderbilt University School of Medicine, Nashville, TN, USA.
authors.forename: Richard M
authors.initials: RM
authors.lastname: Caprioli
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism; D008055:Lipids / Q000235:genetics / Q000302:isolation & purification*; D057054:Molecular Imaging / Q000379:methods*; D011506:Proteins / Q000235:genetics / Q000302:isolation & purification*; D040901:Proteomics; D051381:Rats; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D008055:Lipids; D011506:Proteins
keywords: Cystic fibrosis; FTICR MS; High-throughput; Human clear cell renal cell carcinoma; Nutritional immunity; Technology