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Methods Mol Biol. 2010;664:73-80. doi: 10.1007/978-1-60761-806-5_8.

Tissue microarrays from frozen tissues-OCT technique.

Author information

1
Division of Hematology/Oncology, Department of Medicine, School of Medicine, University of California, Los Angeles, Los Angeles, CA, USA. mfejzo@mednet.ucla.edu

Abstract

Tissue microarray technology is a new method used to analyze 100s to 1,000s of tumor samples on a single slide allowing high throughput analysis of genes and proteins on a large cohort. The original methodology involves coring tissues from paraffin-embedded tissue donor blocks and placing them into a single paraffin block. One difficulty with paraffin-embedded tissue relates to antigenic changes in proteins and mRNA degradation induced by the fixation and embedding process. We modified this technology by using frozen tissues embedded in OCT compound as donor samples and arraying the specimens into a recipient OCT block. Tumor tissue is not fixed before embedding, and sections from the array are evaluated without fixation or postfixed according to the appropriate methodology used to analyze a specific gene at the DNA, RNA, and/or protein levels. While paraffin tissue arrays can be problematic for immunohistochemistry and for RNA in situ hybridization analyses, this method allows optimal evaluation by each technique and uniform fixation across the array panel. Frozen tissue microarrays work well for DNA, RNA, and protein analyses, and may have significant advantages over the original technology for the assessment of some genes and proteins by improving both qualitative and quantitative results.

PMID:
20690054
DOI:
10.1007/978-1-60761-806-5_8
[Indexed for MEDLINE]

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