Cargando…

Luminex-based quantification of Alzheimer’s Disease neuropathologic change in formalin-fixed post-mortem human brain tissue

The vast majority of archived research and clinical pathological specimens are stored in the form of formalin fixed, paraffin-embedded (FFPE) tissues, but, unlike fresh frozen tissue samples, highly quantitative measures in FFPE tissues are limited to immunohistochemical and immunofluorescence thres...

Descripción completa

Detalles Bibliográficos
Autores principales: Keene, C. Dirk, Wilson, Angela M., Kilgore, Mitchell D., Bruner, Lauren T., Postupna, Nadia O., Darvas, Martin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6586549/
https://www.ncbi.nlm.nih.gov/pubmed/30573871
http://dx.doi.org/10.1038/s41374-018-0165-x
_version_ 1783428900267753472
author Keene, C. Dirk
Wilson, Angela M.
Kilgore, Mitchell D.
Bruner, Lauren T.
Postupna, Nadia O.
Darvas, Martin
author_facet Keene, C. Dirk
Wilson, Angela M.
Kilgore, Mitchell D.
Bruner, Lauren T.
Postupna, Nadia O.
Darvas, Martin
author_sort Keene, C. Dirk
collection PubMed
description The vast majority of archived research and clinical pathological specimens are stored in the form of formalin fixed, paraffin-embedded (FFPE) tissues, but, unlike fresh frozen tissue samples, highly quantitative measures in FFPE tissues are limited to immunohistochemical and immunofluorescence thresholding image analysis studies, cell counting, and ordinal ranking systems. This poses a significant obstacle for clinical investigations that aim to correlate diagnostic markers of neurodegenerative diseases like Alzheimer’s disease (AD) with parameters like age, gender, drug exposures, genotype, disease stage, co-morbidities or environmental factors. To overcome this limitation, we have developed Luminex-based techniques and protocols for the quantification of amyloid β and hyperphosphorylated Tau in FFPE brain sections. We validated the Luminex assay in FFPE sections from prefrontal cortex, hippocampus and neostriatum from 30 cases that underwent prior neuropathological diagnostic assessment of AD following the current NIA-AA recommendations for AD: 10 cases diagnosed as not or low, 10 cases as intermediate, and 10 cases as high AD neuropathologic change. Consistent with the neuropathologic assessment, Luminex assay detected high amounts of amyloid beta in the frontal cortex and striatum, and high amounts of hyperphosphorylated Tau in the frontal cortex and hippocampus, of cases with high AD neuropathologic change. This assay can be expanded to detect diverse antigenic targets of interest, as we show here with IBA1 and GFAP. This novel approach supports multiplexed highly quantitative, molecularly specific neuropathology measures to further explore mechanisms of neurodegeneration in AD.
format Online
Article
Text
id pubmed-6586549
institution National Center for Biotechnology Information
language English
publishDate 2018
record_format MEDLINE/PubMed
spelling pubmed-65865492019-06-21 Luminex-based quantification of Alzheimer’s Disease neuropathologic change in formalin-fixed post-mortem human brain tissue Keene, C. Dirk Wilson, Angela M. Kilgore, Mitchell D. Bruner, Lauren T. Postupna, Nadia O. Darvas, Martin Lab Invest Article The vast majority of archived research and clinical pathological specimens are stored in the form of formalin fixed, paraffin-embedded (FFPE) tissues, but, unlike fresh frozen tissue samples, highly quantitative measures in FFPE tissues are limited to immunohistochemical and immunofluorescence thresholding image analysis studies, cell counting, and ordinal ranking systems. This poses a significant obstacle for clinical investigations that aim to correlate diagnostic markers of neurodegenerative diseases like Alzheimer’s disease (AD) with parameters like age, gender, drug exposures, genotype, disease stage, co-morbidities or environmental factors. To overcome this limitation, we have developed Luminex-based techniques and protocols for the quantification of amyloid β and hyperphosphorylated Tau in FFPE brain sections. We validated the Luminex assay in FFPE sections from prefrontal cortex, hippocampus and neostriatum from 30 cases that underwent prior neuropathological diagnostic assessment of AD following the current NIA-AA recommendations for AD: 10 cases diagnosed as not or low, 10 cases as intermediate, and 10 cases as high AD neuropathologic change. Consistent with the neuropathologic assessment, Luminex assay detected high amounts of amyloid beta in the frontal cortex and striatum, and high amounts of hyperphosphorylated Tau in the frontal cortex and hippocampus, of cases with high AD neuropathologic change. This assay can be expanded to detect diverse antigenic targets of interest, as we show here with IBA1 and GFAP. This novel approach supports multiplexed highly quantitative, molecularly specific neuropathology measures to further explore mechanisms of neurodegeneration in AD. 2018-12-20 2019-07 /pmc/articles/PMC6586549/ /pubmed/30573871 http://dx.doi.org/10.1038/s41374-018-0165-x Text en Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:http://www.nature.com/authors/editorial_policies/license.html#terms
spellingShingle Article
Keene, C. Dirk
Wilson, Angela M.
Kilgore, Mitchell D.
Bruner, Lauren T.
Postupna, Nadia O.
Darvas, Martin
Luminex-based quantification of Alzheimer’s Disease neuropathologic change in formalin-fixed post-mortem human brain tissue
title Luminex-based quantification of Alzheimer’s Disease neuropathologic change in formalin-fixed post-mortem human brain tissue
title_full Luminex-based quantification of Alzheimer’s Disease neuropathologic change in formalin-fixed post-mortem human brain tissue
title_fullStr Luminex-based quantification of Alzheimer’s Disease neuropathologic change in formalin-fixed post-mortem human brain tissue
title_full_unstemmed Luminex-based quantification of Alzheimer’s Disease neuropathologic change in formalin-fixed post-mortem human brain tissue
title_short Luminex-based quantification of Alzheimer’s Disease neuropathologic change in formalin-fixed post-mortem human brain tissue
title_sort luminex-based quantification of alzheimer’s disease neuropathologic change in formalin-fixed post-mortem human brain tissue
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6586549/
https://www.ncbi.nlm.nih.gov/pubmed/30573871
http://dx.doi.org/10.1038/s41374-018-0165-x
work_keys_str_mv AT keenecdirk luminexbasedquantificationofalzheimersdiseaseneuropathologicchangeinformalinfixedpostmortemhumanbraintissue
AT wilsonangelam luminexbasedquantificationofalzheimersdiseaseneuropathologicchangeinformalinfixedpostmortemhumanbraintissue
AT kilgoremitchelld luminexbasedquantificationofalzheimersdiseaseneuropathologicchangeinformalinfixedpostmortemhumanbraintissue
AT brunerlaurent luminexbasedquantificationofalzheimersdiseaseneuropathologicchangeinformalinfixedpostmortemhumanbraintissue
AT postupnanadiao luminexbasedquantificationofalzheimersdiseaseneuropathologicchangeinformalinfixedpostmortemhumanbraintissue
AT darvasmartin luminexbasedquantificationofalzheimersdiseaseneuropathologicchangeinformalinfixedpostmortemhumanbraintissue