Frozen tumor tissue microarray technology for analysis of tumor RNA, DNA, and proteins.
Tissue microarray technology is a new method used to analyze several hundred 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 have 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. We show OCT arrays 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.
Schoenberg Fejzo
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Division of Hematology/Oncology, Department of Medicine, University of California, Los Angeles, School of Medicine, Los Angeles, California, USA. mfejzo@mednet.ucla.edu
Slamon
D J
DJ
D000818:Animals; D001943:Breast Neoplasms / Q000737:chemistry; D004273:DNA, Neoplasm / Q000032:analysis*; D005260:Female; D005629:Frozen Sections; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D017404:In Situ Hybridization, Fluorescence; D008175:Lung Neoplasms / Q000737:chemistry; D051379:Mice; D009363:Neoplasm Proteins / Q000032:analysis*; D009368:Neoplasm Transplantation; D009369:Neoplasms / Q000737:chemistry*; D020411:Oligonucleotide Array Sequence Analysis; D012334:RNA, Neoplasm / Q000032:analysis*; D014183:Transplantation, Heterologous; D014407:Tumor Cells, Cultured
D004273:DNA, Neoplasm; D009363:Neoplasm Proteins; D012334:RNA, Neoplasm