Document Detail


Laser capture microdissection for protein and NanoString RNA analysis.
MedLine Citation:
PMID:  23027006     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Yelena Golubeva; Rosalba Salcedo; Claudius Mueller; Lance A Liotta; Virginia Espina
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  931     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2013  
Date Detail:
Created Date:  2012-10-02     Completed Date:  2013-02-11     Revised Date:  2013-09-16    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  213-57     Citation Subset:  IM    
Affiliation:
National Cancer Institute-Frederick/SAIC, Frederick, MD, USA.
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MeSH Terms
Descriptor/Qualifier:
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:
HHSN261200800001E//PHS HHS; R21 CA125698/CA/NCI NIH HHS; R33 CA157403/CA/NCI NIH HHS
Chemical
Reg. No./Substance:
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
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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