title: Adaptation of Laser Microdissection Technique for the Study of a Spontaneous Metastatic Mammary Carcinoma Mouse Model by NanoString Technologies.
abstract: 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.
authors.affiliation: Tumor Growth Factor Section, Mouse Cancer Genetics Program, National Cancer Institute, Frederick, MD, 21702, United States of America.
authors.forename: Nadia P
authors.initials: NP
authors.lastname: Castro
authors.affiliation: CCRIFX Bioinformatics Core, National Cancer Institute, Bethesda, MD, 20892, United States of America.
authors.forename: Anand S
authors.initials: AS
authors.lastname: Merchant
authors.affiliation: Tumor Growth Factor Section, Mouse Cancer Genetics Program, National Cancer Institute, Frederick, MD, 21702, United States of America.
authors.forename: Karen L
authors.initials: KL
authors.lastname: Saylor
authors.affiliation: Pathology-Histotechnology Laboratory, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, Frederick, MD, 21702, United States of America.
authors.forename: Miriam R
authors.initials: MR
authors.lastname: Anver
authors.affiliation: Tumor Growth Factor Section, Mouse Cancer Genetics Program, National Cancer Institute, Frederick, MD, 21702, United States of America.
authors.forename: David S
authors.initials: DS
authors.lastname: Salomon
authors.affiliation: Pathology-Histotechnology Laboratory, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, Frederick, MD, 21702, United States of America.
authors.forename: Yelena G
authors.initials: YG
authors.lastname: Golubeva
mesh_terms: D000818:Animals; D001483:Base Sequence; D045744:Cell Line, Tumor; D005629:Frozen Sections / Q000379:methods; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D061346:Laser Capture Microdissection / Q000379:methods*; D008175:Lung Neoplasms / Q000235:genetics / Q000556:secondary*; D015674:Mammary Neoplasms, Animal / Q000235:genetics / Q000473:pathology*; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008969:Molecular Sequence Data; D036103:Nanotechnology / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis; D020871:RNA Stability; D015203:Reproducibility of Results