In Situ Imaging of Tryptic Peptides by MALDI Imaging Mass Spectrometry Using Fresh‐Frozen or Formalin‐Fixed, Paraffin‐Embedded Tissue. (16th August 2018)

• Record Type: Journal Article
• Authors: Angel, Peggi M.; Norris‐Caneda, Kim; Drake, Richard R.
• Editors: Coligan, John E.; Dunn, Ben M.; Speicher, David W.; Wingfield, Paul T.
• Is Part Of: Current protocols in protein science. Volume 94(2018)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2018-08-16
• Subjects: formalin‐fixed -- imaging mass spectrometry -- MALDI imaging mass spectrometry -- paraffin‐embedded tissue imaging -- peptide identification for imaging mass spectrometry -- proteomics -- tryptic peptide imaging -- tissue imaging; Proteins -- Laboratory manuals; Proteins; Clinical Laboratory Techniques; Genetic Techniques; Immunologic Techniques; Proteins; Laboratory manuals; 572.6028
• DOI: 10.1002/cpps.65
• Languages: English
• ISSNs: 1934-3655

Abstract

Abstract: Tryptic peptide imaging is a primary workflow for matrix‐assisted laser desorption/ionization imaging mass spectrometry (MALDI IMS) and has led to new information reporting highly multiplexed protein localization. Technological advances within the last few years have produced robust tools for automated spraying of both matrix and enzymes. When combined with high‐mass‐resolution and high‐mass‐accuracy instrumentation, studies now generally result in two‐dimensional mapping of well over 1, 000 peptide peaks. This protocol describes sample preparation, spraying, and application of enzymes and matrices, and MALDI FT‐ICR instrumental considerations for two‐dimensional mapping of tryptic peptides from fresh‐frozen or formalin‐fixed, paraffin‐embedded tissue sections. Procedures for extraction of tryptic peptides from tissue sections for LC‐MS/MS identification are also described. © 2018 by John Wiley & Sons, Inc.