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Adaptation of Laser Microdissection Technique for the Study of a Spontaneous Metastatic Mammary Carcinoma Mouse Model by NanoString Technologies

Laser capture microdissection (LCM) of tissue is an established tool in medical research for collection of distinguished cell populations under direct microscopic visualization for molecular analysis. LCM samples have been successfully analyzed in a number of genomic and proteomic downstream molecul...

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Autores principales: Castro, Nadia P., Merchant, Anand S., Saylor, Karen L., Anver, Miriam R., Salomon, David S., Golubeva, Yelena G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4831786/
https://www.ncbi.nlm.nih.gov/pubmed/27077656
http://dx.doi.org/10.1371/journal.pone.0153270
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author Castro, Nadia P.
Merchant, Anand S.
Saylor, Karen L.
Anver, Miriam R.
Salomon, David S.
Golubeva, Yelena G.
author_facet Castro, Nadia P.
Merchant, Anand S.
Saylor, Karen L.
Anver, Miriam R.
Salomon, David S.
Golubeva, Yelena G.
author_sort Castro, Nadia P.
collection PubMed
description Laser capture microdissection (LCM) of tissue is an established tool in medical research for collection of distinguished cell populations under direct microscopic visualization for molecular analysis. LCM samples have been successfully analyzed in a number of genomic and proteomic downstream molecular applications. However, LCM sample collection and preparation procedure has to be adapted to each downstream analysis platform. In this present manuscript we describe in detail the adaptation of LCM methodology for the collection and preparation of fresh frozen samples for NanoString analysis based on a study of a model of mouse mammary gland carcinoma and its lung metastasis. Our adaptation of LCM sample preparation and workflow to the requirements of the NanoString platform allowed acquiring samples with high RNA quality. The NanoString analysis of such samples provided sensitive detection of genes of interest and their associated molecular pathways. NanoString is a reliable gene expression analysis platform that can be effectively coupled with LCM.
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spelling pubmed-48317862016-04-22 Adaptation of Laser Microdissection Technique for the Study of a Spontaneous Metastatic Mammary Carcinoma Mouse Model by NanoString Technologies Castro, Nadia P. Merchant, Anand S. Saylor, Karen L. Anver, Miriam R. Salomon, David S. Golubeva, Yelena G. PLoS One Research Article Laser capture microdissection (LCM) of tissue is an established tool in medical research for collection of distinguished cell populations under direct microscopic visualization for molecular analysis. LCM samples have been successfully analyzed in a number of genomic and proteomic downstream molecular applications. However, LCM sample collection and preparation procedure has to be adapted to each downstream analysis platform. In this present manuscript we describe in detail the adaptation of LCM methodology for the collection and preparation of fresh frozen samples for NanoString analysis based on a study of a model of mouse mammary gland carcinoma and its lung metastasis. Our adaptation of LCM sample preparation and workflow to the requirements of the NanoString platform allowed acquiring samples with high RNA quality. The NanoString analysis of such samples provided sensitive detection of genes of interest and their associated molecular pathways. NanoString is a reliable gene expression analysis platform that can be effectively coupled with LCM. Public Library of Science 2016-04-14 /pmc/articles/PMC4831786/ /pubmed/27077656 http://dx.doi.org/10.1371/journal.pone.0153270 Text en https://creativecommons.org/publicdomain/zero/1.0/ This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 (https://creativecommons.org/publicdomain/zero/1.0/) public domain dedication.
spellingShingle Research Article
Castro, Nadia P.
Merchant, Anand S.
Saylor, Karen L.
Anver, Miriam R.
Salomon, David S.
Golubeva, Yelena G.
Adaptation of Laser Microdissection Technique for the Study of a Spontaneous Metastatic Mammary Carcinoma Mouse Model by NanoString Technologies
title Adaptation of Laser Microdissection Technique for the Study of a Spontaneous Metastatic Mammary Carcinoma Mouse Model by NanoString Technologies
title_full Adaptation of Laser Microdissection Technique for the Study of a Spontaneous Metastatic Mammary Carcinoma Mouse Model by NanoString Technologies
title_fullStr Adaptation of Laser Microdissection Technique for the Study of a Spontaneous Metastatic Mammary Carcinoma Mouse Model by NanoString Technologies
title_full_unstemmed Adaptation of Laser Microdissection Technique for the Study of a Spontaneous Metastatic Mammary Carcinoma Mouse Model by NanoString Technologies
title_short Adaptation of Laser Microdissection Technique for the Study of a Spontaneous Metastatic Mammary Carcinoma Mouse Model by NanoString Technologies
title_sort adaptation of laser microdissection technique for the study of a spontaneous metastatic mammary carcinoma mouse model by nanostring technologies
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4831786/
https://www.ncbi.nlm.nih.gov/pubmed/27077656
http://dx.doi.org/10.1371/journal.pone.0153270
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