High-Throughput Microdissection for Next-Generation Sequencing: e0151775

• Published in: PloS one Vol. 11; no. 3
• Main Authors: Rosenberg, Avi Z, Armani, Michael D, Fetsch, Patricia A, Xi, Liqiang, Pham, Tina Thu, Raffeld, Mark, Chen, Yun, O'Flaherty, Neil, Stussman, Rebecca, Blackler, Adele R
• Format: Journal Article
• Language: English
• Published: 01.03.2016
• Subjects: Epstein-Barr virus
• ISSN: 1932-6203
• DOI: 10.1371/journal.pone.0151775

Precision medicine promises to enhance patient treatment through the use of emerging molecular technologies, including genomics, transcriptomics, and proteomics. However, current tools in surgical pathology lack the capability to efficiently isolate specific cell populations in complex tissues/tumors, which can confound molecular results. Expression microdissection (xMD) is an immuno-based cell/subcellular isolation tool that procures targets of interest from a cytological or histological specimen. In this study, we demonstrate the accuracy and precision of xMD by rapidly isolating immunostained targets, including cytokeratin AE1/AE3, p53, and estrogen receptor (ER) positive cells and nuclei from tissue sections. Other targets procured included green fluorescent protein (GFP) expressing fibroblasts, in situ hybridization positive Epstein-Barr virus nuclei, and silver stained fungi. In order to assess the effect on molecular data, xMD was utilized to isolate specific targets from a mixed population of cells where the targets constituted only 5% of the sample. Target enrichment from this admixed cell population prior to next-generation sequencing (NGS) produced a minimum 13-fold increase in mutation allele frequency detection. These data suggest a role for xMD in a wide range of molecular pathology studies, as well as in the clinical workflow for samples where tumor cell enrichment is needed, or for those with a relative paucity of target cells.