Laser microdissection

Lehmann U, Bock O, Glockner S, et al. Quantitative molecular analysis of laser-microdissected paraffin-embedded human tissues. Pathobiology 2000 68 202-208. 

Lahr G, Sticha M, Schtitzea K, et al. Diagnosis of papillary thyroid carcinoma is facilitated by using an RT-PCR approach on laser-microdissected archival material to detect RET oncogene activation. Pathobiology 2000 68 218-226. 

The first step in all RNA isolation protocols involves lysing the cell in a chemical environment that denatures ribonucleases. The RNA is then fractionated from other cellular macromolecules by either homogenizing the tissue (dissected brain tissue) or simply vortexing the sample (very small tissues and laser-microdissected sample) without further homogenization. The cell type from which the RNA is to be isolated, the sample size, and the eventual use of the RNA will determine which procedure described here is appropriate. 

The routine competitive RT-PCR protocol described earlier is modified for the quantitative nested RT-PCR analysis of RNA isolated from laser-microdissected samples. The low amount of starting template and the... 

Vandewoestyne ML, Pede VC, Lambein KY et al (2011) Laser microdissection for the assessment of the clonal relationship between chronic lymphocytic leukemia/smaU lymphocytic lymphoma and proliferating B cells within lymph node pseudofolKcles. Leukemia 25 883-888... 

Kondo T, Seike M, Mori Y, Fujii K, Yamada T, Hirohashi S. (2003) Application of sensitive fluorescent dyes in linkage of laser microdissection and two-dimensional gel electrophoresis as a cancer proteomic study tool. Proteomics 3, 1758-66. 

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. 

Yang F, Foekens JA, Yu J et al. Laser microdissection and microarray analysis of breast tumors reveal ER-alpha-related genes and pathways. Oncogene 2006 25 1413-1419. 

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. 

Sauna PP, King AR, van der Stap LD, Repunte-Canonigo V (2005) Gene profiling of laser-microdissected brain regions and sub-regions. Brain Res Brain Res Pi otoc 15 66-74. 

N. P. Gerry, M.E. Lenburg, C.L. Rosenberg, Reliability and reproducibility of gene expression measurements using amplified RNA from laser-microdissected primary breast tissue with oligonucleotide arrays, J. Mol. Diagn. 2005, 7, 57-64. 

Ogawa K, Asamoto M, Suzuki S, et al. (2005). Downregulation of apoptosis revealed by laser microdissection and cDNA microarray analysis of related genes in rat liver preneoplastic lesions. Medical Molec. Morphol. 38 23-29. 

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. 

Greengauz-Roberts, O., Sloppier, H Nomura, S., Yamaguchi, H Goldenring, J. R., Podolsky, R. H., Lee, J. R., and Dynan, W.S. (2005) Saturation labeling with cysteine-reactive cyanine fluorescent dyes provides increased sensitivity for protein expression profiling of laser-microdissected clinical specimens. 

Umar, A. Luider, T.M. Foekens, J.A. Pasa-Tolic, L. Nano-LC FT-ICR MS improves proteome coverage attainable for similar to 3000 laser-microdissected breast carcinoma cells. Proteomics 2007, 7(2), 323-329. 

Bolster, J., Dithmar, H., Bmgemeister, R., Friedemann, G. Feucht, W. (2006). Flavonoids in plant nuclei detection by laser microdissection and pressure catapulting (LMPC), in vivo staining, and UV-visible spectroscopic titration. Physiologia Plantarum, 128, 163-174. 

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. 

Vega, C. J. Laser microdissection sample preparation for RNA analyses. Methods Mol. Biol. 2008, 414, 241-252. 

In parallel, many efforts in the proteomic and metabolomic areas were devoted to the development of analytical protocols making it possible to individualize, within biological samples, differentiated compartments. For instance, the histological analysis of a biological tissue section followed by the laser microdissection of relevant regions can be proposed as a preliminary stage to an omic analysis intended to distinguish the chemical content of close tissue zones. In addition, approaches developed for the smdy of well-defined cellular compartments (such as membranes or mitochondria) enable one to differentiate subcellular... 

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