Laser-captured microdissection

Torres-Munoz JE, Nunez M, Petito CK (2008) Successful Application of Hyperbranched Multidisplacement Genomic Amplification to Detect HlV-1 Sequences in Single Neurons Removed from Autopsy Brain Sections by Laser Capture Microdissection. J Mol Diagn 10(4) 317-324... 

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

Numerous articles have demonstrated the availability of RNA extracted from a few cells or even a single cell taken by laser captured microdissection (LCM) system from archival FFPE tissue sections that had been previously stained by IFIC. Using a combination of pre-immunostained FFPE tissue section with LCM, a sensitive real-time quantitative RT-PCR can be achieved based on a few immuno-detected cells, creating a way to study pathophysiological gene regulation in a cell-specific manner in archival tissues housed in... 

Jin L, Majerus J, Oliveira A, et al. Detection of fusion gene transcripts in fresh-frozen and formalin-fixed paraffin-embedded tissue sections of soft-tissue sarcomas after laser capture microdissection and RT-PCR. Diagn. Mol. Pathol. 2003 12 224-230. 

Lindeman N, Waltregny D, Signoretti S, et al. Gene transcript quantitation by real-time RT-PCR in cells selected by immunohistochemistry-laser capture microdissection. Diagn. Mol. Pathol. 2002 11 187-192. 

Tanji N, Ross MD, Cara A, et al. Effect of tissue processing on the ability to recover nucleic acid from specific renal tissue compartments by laser capture microdissection. Exp. Nephrol. 2001 9 229-234. 

EVALUATION OF LASER CAPTURE MICRODISSECTION OF FFPE TISSUES... 

LCM, laser captured microdissection ESI, electrospray ionization PAGE, polyacrylamide gel electrophoresis IGF-1R, insulin-like growth factor receptor 1. 

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. 

RNA Isolation from Laser-Capture Microdissected Samples. 355... 

During recent years, many of our appheations of competitive RT-PCR have involved the analysis of RNA from very small tissue samples obtained by laser capture microdissection. We have therefore adopted a modified reamphfication protocol (competitive Nested-RT-PCR) for the quantitation of very small tissue samples (Costa et al., 2002). The procedure involves the use of very low concentrations of cRNA (internal standard) in the presence of total RNA isolated from a measured and microdissected tissue sample or a... 

Quantitation of RNA Transcripts from Laser-Capture Microdissection Material... 

Luzzi V, Holtschlag V, Watson MA. 2001. Expression profiling of ductal carcinoma in situ by laser capture microdissection and high density oligonucleotides arrays. Am J Pathol 158 2005. 

Batorfi, J., Ye, B., Mok, S.C., Cseh, I., Berkowitz, R.S., and Fulop, V., Protein profiling of complete mole and normal placenta using ProteinChip analysis of laser capture microdissected cells, Gynecol. Oncol., 88, 424-428, 2003. 

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... 

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... 

Cell Migration Developmental Methods and Protocols, edited by Jun-Lin Guan, 2005 293. Laser Capture Microdissection Methods and Protocols, edited by Graeme I. Murray and Stephanie Curran, 2005... 

Emmert-Buck MR, Bonner RE, Smith PD, et al. (1996) Laser capture microdissection. Science 274, 998-1001. 

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. 

Craven BA, Totty N, Harnden P, Selby PJ, Banks RE. (2002) Laser capture microdissection and two-dimensional polyacrylamide gel electrophoresis evaluation of tissue preparation and sample limitations. Am J Pathol 160, 815-22. 

Microdissection of a tumor was initially carried out using a standard syringe (5). The tumor was viewed under a microscope so that it could be separated from the surrounding normal tissues. For this method to be effective, tumors needed to be easily defined under the microscope, which limited the number of samples that were compatible. Microdissection techniques have further evolved to include techniques such as laser microbeam microdissection (EMM) and laser capture microdissection (LCM). These... 

While laser capture microdissection and other related techniques may allow for the generation of relatively homogenous cell populations, most such purification steps will largely eliminate any potential cancer stem cells, because these techniques necessitate the identification of differentiated cell types for isolation. As such, cancer stem cells would frequently be eliminated from analysis, and any cancer stem cells that may be accidentally included in the analysis would be in such low occurrence that their signature would most certainly be missed. 

Suarez-Quian CA, Goldstein SR, Pohida T et al. Laser capture microdissection of single cells from complex tissues. Biotechniques 1999 26 328-335. 

Fend F, Raffeld M. Laser capture microdissection in pathology. J Clin Pathol 2000 53 666-672. Alevizos I, Mahadevappa M, Zhang X et al. Oral cancer in vivo gene expression profiling assisted by laser capture microdissection and microarray analysis. Oncogene 2001 20 6196-6204. 

Ohyama H, Zhang X, Kohno Y et al. Laser capture microdissection-generated target sample for high-density oligonucleotide array hybridization. Biotechniques 2000 29 530-536. 

Wu MS, Lin YS, Chang YT et al. Gene expression profiling of gastric cancer by microarray combined with laser capture microdissection. World J Gastroenterol 2005 11 7405-7412. 

---