Laser capture microdissection proteomics

Ornstein DK et al. Proteomic analysis of laser capture microdissected human prostate cancer and in vitro prostate cell lines. Electrophoresis 2000 21 2235-2242. 

The sample materials from which proteins for proteomics studies may be extracted include fresh or snap-frozen cells from varied sources such as biological fluids, (serum, urine, plasma) and solid tissues such as biopsy specimens. Moreover, proteins isolated from ethanol-fixed paraffin-embedded tissues can be utilized for MS analysis.2 Protocols for the identification of proteins from formalin-fixed paraffin-embedded (FFPE) tissues have been recently developed.3 4 FFPE materials are the most common forms of biopsy archives utilized worldwide, and represent an important advancement for the large-scale interrogation of proteins in archival patient-derived materials. Finally, laser capture microdissected tissues have been successfully used for MS analysis.45... 

An alternative approach to assessing tissue-specific expression at the proteomic level can be achieved by MS of laser capture microdissected tissues.4 An important development in this arena is the ability to perform LCM and MS/MS on formalin-fixed paraffin-embedded tissues. 

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... 

VanMeter AJ, Rodriguez AS, Bowman ED et al (2008) Laser capture microdissection and protein microarray analysis of human non-smaU cell lung cancer differential epidermal growth factor receptor (EGPR) phosphorylation events associated with mutated EGFR compared with wild type. Mol Cell Proteomics 7 1902-1924... 

Banks BE, Dunn MJ, Eorbes MA, et al. (1999) The potential use of laser capture microdissection to selectively obtain distinct populations of ceUs for proteomic analysis - preliminary findings. Electrophoresis 20, 689-700. 

Li C, Hong Y, Tan YX et al. Accurate qualitative and quantitative proteomic analysis of clinical hepatocellular carcinoma using laser capture microdissection coupled with isotope-coded affinity tag and two-dimensional liquid chromatography mass spectrometry. Mo/ Cell Proteomics 2004,3399-409. 

In 2001, both Lawrie et al. and Moskaluk coupled LCM (laser capture microdissection) with 2D gel proteomics to recover proteins from laser captured micro-dissected tissue in a form that can be used in 2D gel analysis and mass spectrometry. This may provide valuable information for protein profiling and databasing of human tissues in healthy and disease states. 

Mouledous L, Hunt S, Harcourt R, Harry J, Williams KL, Gutstein HB. Navigated laser capture microdissection as an alternative to direct histological staining for proteomic analysis of brain samples. Proteomics 2003 3(5) 610-615. 

The importance for proteomic studies of being performed on standardized samples cannot be emphasized enough. Several approaches have been developed to address the problem of tissue heterogeneity. Beads-based sample enrichment (BBSE) and laser capture microdissection (LCM) procedures are emerging as the methods of choice. The aim of these procedures is to eliminate, as much as possible, all confounding and contaminating factors in order to obtain a sample quality that can be compared with cell lines without the disadvantages of in vitro cultures. 

Craven RA, Banks RE. Laser capture microdissection and proteomics Possibilities and limitation. Proteomics 2001 1(10) 1200—1204. Review. 

Lawrie LC, Curran S. Laser capture microdissection and colorectal cancer proteomics. Methods Mol Biol 2005 293 245-253. 

Shekouh AR, Thompson CC, Prime W, Campbell F, Hamlett J, Herrington CS, et al. Application of laser capture microdissection combined with two-dimensional electrophoresis for the discovery of differentially regulated proteins in pancreatic ductal adenocarcinoma. Proteomics 2003 3(10) 1988—2001. 

Gearing, M., Rees, H. D., Iah, J. J., Levey, A. I., Peng, J. (2004). Proteomic characterization of postmortem amyloid plaques isolated by laser capture microdissection. J. Biol. Chem. 279, 37061-37068. 

Use of laser capture microdissection to selectively obtain distinct populations of cells for proteomic analysis. Methods Enzymol. 356, 33—49. 

Mouledous, L., Hunt, S., Harcourt, R., Harry, J.L., Williams, K.L. and Gutstein, H.B. (2003) Proteomic analysis of immunostained, laser-capture microdissected brain samples. Electrophoresis 24,296-302. Nagy, A. and Delgado-Escueta, A.V. (1984) Rapid preparation of synaptosomes from mammalian brain using nontoxic isoosmotic gradient material (Percoll). J. Neurochem. 43, 1114-1123. 

Wittliff JL, Erlander MG. Laser capture microdissection and its applications in genomics and proteomics. Meth Enzymol 2002 356 12-25. 

Jain KK. Application of laser capture microdissection to proteomics. Meth Enzymol 2002 356 157-67. 

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). 

Automated Laser Capture Microdissection for Tissue Proteomics... 

Key Words Cancer DNA laser capture microdissection molecular profiling proteomics protein RNA tissue heterogeneity. 

The fields of genomics and proteomics have been enhanced hy cellular isolation techniques such as Laser Capture Microdissection (LCM), which enables the isolation and molecular investigation of pure cell populations through direct... 

Cowherd, S.M., Espina, V.A., Petricoin, E.F., 3rd, and Liotta, L.A. (2004) Proteomic analysis of human breast cancer tissue with laser-capture microdissection and reverse-phase protein microarrays. Clin Breast Cancer 5, 385-92. 

Zang, L., Palmer Toy, D., Hancock, W.S., Sgroi, D.C., and Karger, B.L. (2004) Proteomic analysis of ductal carcinoma of the breast using laser capture microdissection, LC-MS, and 160/180 isotopic labeling. J Proteome Res 3, 604-12. 

Key Words Cancer laser capture microdissection microarray molecnlar profiling protein proteomics tissue heterogeneity theranostics. 

The fifth chapter in this volume is unique in that it presents a very useful protocol, however, one that must be used in conjunction with other methods. Automated laser capture microdissection, which is used to capture homogeneous cell populations from crude tissue samples may be used in concert with a number of the other platforms presented in this volume. This technique should be considered a valuable tool for researchers in tissue proteomics, especially for the production of custom arrays. 

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