High-Throughput Microdissection for Next-Generation Sequencing

Precision medicine promises to enhance patient treatment through the use of emerging molecular technologies, including genomics, transcriptomics, and proteomics. However, current surgical pathology tools lack the capability to efficiently isolate specific cell populations, which can confound molecular results. Expression microdissection (xMD) is an immuno-based cell/subcellular isolation tool that procures targets of interest from a cytological/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. In addition, procurement of green fluorescent protein (GFP) expressing fibroblasts, in situ hybridization of Epstein-Barr virus nuclei, and silver stained fungi targets was successfully performed. Finally, xMD was utilized to isolate specific targets from a mixed population of cells, where the targets only constituted 5% of the sample. Target enrichment from this heterogeneous cell population prior to next-generation sequencing (NGS) studies produced a minimum 13-fold increase in mutation detection. These data suggest a role for xMD in the clinical workflow, especially for samples with a relative paucity of target cells.