Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
1226311 | Journal of Proteomics | 2012 | 13 Pages |
One of the important challenges for MALDI imaging mass spectrometry (MALDI-IMS) is the unambiguous identification of measured analytes. One way to do this is to match tryptic peptide MALDI-IMS m/z values with LC-MS/MS identified m/z values. Matching using current MALDI-TOF/TOF MS instruments is difficult due to the variability of in situ time-of-flight (TOF) m/z measurements. This variability is currently addressed using external calibration, which limits achievable mass accuracy for MALDI-IMS and makes it difficult to match these data to downstream LC-MS/MS results. To overcome this challenge, the work presented here details a method for internally calibrating data sets generated from tryptic peptide MALDI-IMS on formalin-fixed paraffin-embedded sections of ovarian cancer. By calibrating all spectra to internal peak features the m/z error for matches made between MALDI-IMS m/z values and LC-MS/MS identified peptide m/z values was significantly reduced. This improvement was confirmed by follow up matching of LC-MS/MS spectra to in situ MS/MS spectra from the same m/z peak features. The sum of the data presented here indicates that internal calibrants should be a standard component of tryptic peptide MALDI-IMS experiments.This article is part of a Special Issue entitled: Imaging Mass Spectrometry: A User’s Guide to a New Technique for Biological and Biomedical Research.
Graphical abstractFigure optionsDownload full-size imageDownload high-quality image (193 K)Download as PowerPoint slideHighlights► This study improved MALDI imaging data set mass accuracy using internal calibrants. ► Internal calibration significantly improved matching of MALDI imaging and LC-MS/MS data. ► Improved matches were confirmed by comparing in situ MS/MS to higher quality LC-MS/MS data.