| Laser capture microdissection for protein and NanoString RNA analysis. | |
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MedLine Citation:
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PMID: 23027006 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Laser capture microdissection (LCM) allows the precise procurement of enriched cell populations from a heterogeneous tissue, or live cell culture, under direct microscopic visualization. Histologically enriched cell populations can be procured by harvesting cells of interest directly or isolating specific cells by ablating unwanted cells. The basic components of laser microdissection technology are (a) visualization of cells via light microscopy, (b) transfer of laser energy to a thermolabile polymer with either the formation of a polymer-cell composite (capture method) or transfer of laser energy via an ultraviolet laser to photovolatize a region of tissue (cutting method), and (c) removal of cells of interest from the heterogeneous tissue section. The capture and cutting methods (instruments) for laser microdissection differ in the manner by which cells of interest are removed from the heterogeneous sample. Laser energy in the capture method is infrared (810 nm), while in the cutting mode the laser is ultraviolet (355 nm). Infrared lasers melt a thermolabile polymer that adheres to the cells of interest, whereas ultraviolet lasers ablate cells for either removal of unwanted cells or excision of a defined area of cells. LCM technology is applicable to an array of applications including mass spectrometry, DNA genotyping and loss-of-heterozygosity analysis, RNA transcript profiling, cDNA library generation, proteomics discovery, and signal kinase pathway profiling. This chapter describes LCM using an Arcturus(XT) instrument for downstream protein sample analysis and using an mmi CellCut Plus® instrument for RNA analysis via NanoString technology. |
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Authors:
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Yelena Golubeva; Rosalba Salcedo; Claudius Mueller; Lance A Liotta; Virginia Espina |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural |
Journal Detail:
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Title: Methods in molecular biology (Clifton, N.J.) Volume: 931 ISSN: 1940-6029 ISO Abbreviation: Methods Mol. Biol. Publication Date: 2013 |
Date Detail:
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Created Date: 2012-10-02 Completed Date: 2013-02-11 Revised Date: 2013-05-08 |
Medline Journal Info:
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Nlm Unique ID: 9214969 Medline TA: Methods Mol Biol Country: United States |
Other Details:
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Languages: eng Pagination: 213-57 Citation Subset: IM |
Affiliation:
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National Cancer Institute-Frederick/SAIC, Frederick, MD, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Base Sequence Coloring Agents / chemistry Cryopreservation Ear, Inner / cytology Eosine Yellowish-(YS) / chemistry Gene Expression Profiling* Hematoxylin / chemistry Laser Capture Microdissection / methods* Mice Microtomy Molecular Sequence Data Oxazines / chemistry Papilloma / pathology Paraffin Embedding Proteins / genetics, metabolism* RNA / genetics, metabolism* Skin Neoplasms / pathology Staining and Labeling |
| Grant Support | |
ID/Acronym/Agency:
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HHSN261200800001E//PHS HHS; R21 CA125698/CA/NCI NIH HHS; R33 CA157403/CA/NCI NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Coloring Agents; 0/Oxazines; 0/Proteins; 17372-87-1/Eosine Yellowish-(YS); 18472-89-4/cresyl violet; 517-28-2/Hematoxylin; 63231-63-0/RNA |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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