z-logo
Premium
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
Author(s) -
Judd Audra M.,
Gutierrez Danielle B.,
Moore Jessica L.,
Patterson Nathan Heath,
Yang Junhai,
Romer Carrie E.,
Norris Jeremy L.,
Caprioli Richard M.
Publication year - 2019
Publication title -
journal of mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 121
eISSN - 1096-9888
pISSN - 1076-5174
DOI - 10.1002/jms.4384
Subject(s) - chemistry , mass spectrometry imaging , in situ , mass spectrometry , maldi imaging , sample preparation , matrix assisted laser desorption/ionization , matrix (chemical analysis) , desorption , chromatography , desorption electrospray ionization , native tissue , computational biology , analytical chemistry (journal) , biomedical engineering , ionization , tissue engineering , chemical ionization , ion , organic chemistry , adsorption , biology , medicine
Abstract 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.

This content is not available in your region!

Continue researching here.

Having issues? You can contact us here