Laser technologies 553 microdissection

The term reverse phase protein microarray was first coined in 2001 by Paweletz et al. (5) in the landmark paper which introduced the platform and demonstrated its utility for analyzing proteins derived from laser capture microdissected (LCM) prostate cancer. RPMAs have also been referred to by other names in the literature including lysate arrays, reverse phase lysate arrays, protein microarrays, and reverse phase protein arrays (6). A very nice historical perspective on the development of RPJMA technology is presented in a review by Mueller et al. (6). A detailed description of the methods for performing RPMA is beyond the scope of this chapter and would be redundant because there are... 

Paweletz CP, Liotta LA, Petricorn EE (2001) New technologies for biomarker analysis of prostate cancer progression laser capture microdissection and tissue proteomics. Urology 57 160-163... 

Paweletz, C.P Liotta, L.A. Petricoin, E.F. New Technologies for Biomarker Analysis of Prostate Cancer Progression Laser Capture Microdissection and Tissue Proteomics, Urology 57,160-163 (2001). 

Laser capture microdissection for obtaining pure cells from tissues rmder direct vision Display technologies fluorescence and chemiluminescence ... 

Laser capture microdissection (LCM) provides an ideal method for extraction of cells from specimens in which the exact morphologies of both the captured cells and the surrounding tissue are preserved. The following proteomics technologies, which are relevant to drug discovery, can be combined with LCM ... 

Melle, C., Ernst, G., Schimmel, B., Bleul, A, Koscielny, S., Wiesner, A., Bogumil, R., Moller, U., Osterloh, D., Halbhuber, K.J., and F. Von Eggling, 2003, Biomarker Discovery and Identification in Laser Microdissected Head and Neck Squamous Cell Carcinoma with ProteinChip(R) Technology, Two-dimensional Gel Electrophoresis, Tandem Mass Spectrometry, and Immunohistochemistry. Mol Cell Proteomics. 2(7) 443-52. 

The primary components of LCM technology are (1) visualization of the cells of interest through microscopy, (2) transfer of near-infrared laser energy pulses to a thermolabile polymer with formation of a polymer-cell composite, and (3) removal of the polymer from the tissue surface, which shears the embedded cells of interest away from the heterogeneous tissue section (18,19). Extraction buffers applied to the polymer film solubilize the cells, liberating the molecules of interest. The DNA, RNA, or protein from the microdissected cells may be analyzed by any method with appropriate sensitivity (20,21,22,23,24). Protein extracted from microdissected cells may be used for mass spectrometric analysis, applied to reverse phase protein microarrays, or used for western blot analysis (25,26). 

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