Laser Capture Microdissection for Protein and NanoString RNA Analysis View Full Text


Ontology type: schema:Chapter      Open Access: True


Chapter Info

DATE

2012

AUTHORS

Yelena Golubeva , Rosalba Salcedo , Claudius Mueller , Lance A. Liotta , Virginia Espina

ABSTRACT

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. More... »

PAGES

213-257

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/978-1-62703-056-4_12

DOI

http://dx.doi.org/10.1007/978-1-62703-056-4_12

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1028302488

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/23027006


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