Proteomic analysis microdissection

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

Guo T, Wang W, Rudnick PA, et al. Proteome analysis of microdissected formalin-fixed and paraffin-embedded tissue specimens. J. Histochem. Cytochem. 2007 55 763-772. 

SHOTGUN PROTEOME ANALYSIS OF MICRODISSECTED FORMALIN-FIXED BRAIN TUMOR TISSUE... 

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. 

Wilson KE, Marouga R, Prime JE, et al. (2005) Comparative proteomic analysis using samples obtained with laser microdissection and saturation dye labelling. Proteomics 5, 3851-8. 

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. 

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. 

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. 

Wilson, K. E., Marouga, R., Prime, J. E D. Pashby, P., Orange, P. R., Crosier, S., Keith, A. B Lathe, R., Mullins, J., Estibeiro, P., Bergling, H., Hawkins, E and Morris, C. M. (2005) Comparative proteomic analysis using samples obtained with laser microdissection and saturation dye labeling. Proteomics 5, 3851-58. 

Optimal tissue thickness for microdissection is 5-8 pm. Tissue sections less than 5 pm may not provide a full cell thickness, necessitating microdissection of more cells for a given assay. Tissue sections thicker than 8 pm may not microdissect completely, leaving integral cellular components adhering to the slide. Ethanol fixed tissue sections may also be used for proteomic analysis, but a larger number of microdissected cells may be required for adequate sensitivity in downstream assays as compared with frozen sections. 

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. 

Banks, R., et al. (1999). The Potential Use of Laser Capture Microdissection to Selectively Obtain Distinct Populations of Cells for Proteomic Analysis Preliminary Findings, Electrophoresis 20 689-700. 

Vrana JA, Gamez JD, Madden BJ, Theis JD, Bergen HR, Dogan A. Classification of amyloidosis by laser microdissection and mass spectrometry-based proteomic analysis in clinical biopsy specimens. Blood 2009 114 4957—9. 

A number of proteomic studies on archival material have utilized Liquid Tissue (Expression Pathology, Inc., Gaithersburg, MD), a commercial protein extraction kit for FFPE tissue.4,9,25-28 This kit is also based upon HIAR techniques and shares a similar work flow to the methods already discussed. Thin, typically 5-10pM, sections are cut from paraffin tissue blocks, the paraffin is removed, and the tissue deparaffinized and rehydrated in alcohols and distilled water before microdissection. The cellular material is then suspended in Liquid Tissue buffer and heated at 95°C for 90 min. Trypsin is added, and the material is digested overnight at 37°C prior to reduction with DTT and analysis by LC-MS/MS.26... 

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... 

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