GOP-10: Post-mortem diagnosis of myocardial ischemia. A feasibility study on the application of LA-ICP MS, with multiplex quantitative imaging
Détails
ID Serval
serval:BIB_42C563D6F571
Type
Actes de conférence (partie): contribution originale à la littérature scientifique, publiée à l'occasion de conférences scientifiques, dans un ouvrage de compte-rendu (proceedings), ou dans l'édition spéciale d'un journal reconnu (conference proceedings).
Sous-type
Abstract (résumé de présentation): article court qui reprend les éléments essentiels présentés à l'occasion d'une conférence scientifique dans un poster ou lors d'une intervention orale.
Collection
Publications
Institution
Titre
GOP-10: Post-mortem diagnosis of myocardial ischemia. A feasibility study on the application of LA-ICP MS, with multiplex quantitative imaging
Titre de la conférence
IALM intersocietal symposium
Adresse
Venise, Italie, 21-24 juin 2016
Statut éditorial
Publié
Date de publication
2016
Peer-reviewed
Oui
Langue
anglais
Résumé
Background and Aims.
Post-mortem diagnosis of myocardial ischemia is known to be challenging for forensic and clinical pathologists, especially if death occurred shortly after the onset of ischemia. Many published studies have shown that immunohistochemistry can considerably improve the diagnosis, especially when not a single marker, but a panel of markers, is used. One of the major limits of conventional immunohistochemistry is the inability to detect multiple antigens in a single tissue section, because of spectral overlap of tags, observed using either chromogenic (maximum 3 colors are available) or fluorescent (4-5 colors are available) detection. Moreover, when performing multiplex immunohistochemistry, if indirect detection methods are employed, cross-reactivity between the detection reagents can take place.
Materials and Methods.
Here we present the use of laser ablation-inductively coupled plasma mass spectrometry (LA-ICP MS) for the multiplex quantitative imaging of markers in tissue section at cellular resolution (i.e. 1 Oμm). In this feasibility study we tested six primary antibodies, chosen among those we currently use in our routine and research activity in order to detect acute myocardial ischemia. These antibodies (troponin T, myoglobin, fibronectin, C5b-9, connexin 43 and JunB) were labeled each with a rare earth metal isotope prior to their simultaneous incubation on tissue section. We compared the LA-ICP MS approach and conventional immunohistochemistry, using five formalin fixed, paraffin embedded (FFPE) autopsy samples of human ischemic myocardium.
Results and Conclusions.
A good correlation was obtained between ionic and optical images, providing similar detection patterns. Furthermore, we determined the absolute concentration of markers from the LA-ICP MS acquisition, and we were able to show a proportional relationship between signal intensity and local marker concentration in thin tissue sections. The LA-ICP MSbased method is a novel approach for multiplex mapping and imaging of antigen-antibody complexes in tissue, with a relevant diagnostic potential for forensic and clinical pathology.
Post-mortem diagnosis of myocardial ischemia is known to be challenging for forensic and clinical pathologists, especially if death occurred shortly after the onset of ischemia. Many published studies have shown that immunohistochemistry can considerably improve the diagnosis, especially when not a single marker, but a panel of markers, is used. One of the major limits of conventional immunohistochemistry is the inability to detect multiple antigens in a single tissue section, because of spectral overlap of tags, observed using either chromogenic (maximum 3 colors are available) or fluorescent (4-5 colors are available) detection. Moreover, when performing multiplex immunohistochemistry, if indirect detection methods are employed, cross-reactivity between the detection reagents can take place.
Materials and Methods.
Here we present the use of laser ablation-inductively coupled plasma mass spectrometry (LA-ICP MS) for the multiplex quantitative imaging of markers in tissue section at cellular resolution (i.e. 1 Oμm). In this feasibility study we tested six primary antibodies, chosen among those we currently use in our routine and research activity in order to detect acute myocardial ischemia. These antibodies (troponin T, myoglobin, fibronectin, C5b-9, connexin 43 and JunB) were labeled each with a rare earth metal isotope prior to their simultaneous incubation on tissue section. We compared the LA-ICP MS approach and conventional immunohistochemistry, using five formalin fixed, paraffin embedded (FFPE) autopsy samples of human ischemic myocardium.
Results and Conclusions.
A good correlation was obtained between ionic and optical images, providing similar detection patterns. Furthermore, we determined the absolute concentration of markers from the LA-ICP MS acquisition, and we were able to show a proportional relationship between signal intensity and local marker concentration in thin tissue sections. The LA-ICP MSbased method is a novel approach for multiplex mapping and imaging of antigen-antibody complexes in tissue, with a relevant diagnostic potential for forensic and clinical pathology.
Mots-clé
Myocardial ischemia, immunohistochemistry, laser ablation-inductively coupled plasma mass spectrometry, multiplex
Création de la notice
13/07/2016 11:47
Dernière modification de la notice
20/08/2019 13:45
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