A recommended and verified procedure for in situ tryptic digestion of formalin-fixed paraffin-embedded tissues for analysis by matrix-assisted laser desorption/ionization imaging mass spectrometry
| Autor: | Richard M. Caprioli, Junhai Yang, Carrie E Romer, Nathan Heath Patterson, Audra M. Judd, Jessica L. Moore, Jeremy L. Norris, Danielle B. Gutierrez |
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| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
In situ
Tissue Fixation Formalin fixed paraffin embedded 01 natural sciences Verified procedure Mass spectrometry imaging Article Specimen Handling Digestion (alchemy) Formaldehyde Humans Sample preparation Trypsin Spectroscopy Chromatography Paraffin Embedding 010405 organic chemistry Chemistry 010401 analytical chemistry Proteins 0104 chemical sciences Matrix-assisted laser desorption/ionization Tissue Array Analysis Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Proteolysis Molecular imaging |
| Zdroj: | J Mass Spectrom |
| Popis: | Matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI IMS) is a molecular imaging technology uniquely capable of untargeted measurement of proteins, lipids, and metabolites while retaining spatial information about their location in situ. This powerful combination of capabilities has the potential to bring a wealth of knowledge to the field of molecular histology. Translation of this innovative research tool into clinical laboratories requires the development of reliable sample preparation protocols for the analysis of proteins from formalin-fixed paraffin embedded (FFPE) tissues, the standard preservation process in clinical pathology. Although ideal for stained tissue analysis by microscopy, the FFPE process cross-links, disrupts, or can remove proteins from the tissue, making analysis of the protein content challenging. To date, reported approaches differ widely in process and efficacy. This tutorial presents a strategy, derived from systematic testing and optimization of key parameters, for reproducible in situ tryptic digestion of proteins in FFPE tissue and subsequent MALDI IMS analysis. The approach describes a generalized method for FFPE tissues originating from virtually any source. |
| Databáze: | OpenAIRE |
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