Molecular analysis of live rare cells, such as circulating tumor cells (CTCs) is challenging. Current strategies include antibody-mediated capture and size selection and are limited in the purity and recovery of the cells of interest. We have developed a novel approach for rare cell analysis based on the specific lysis of targeted cells in a background of a vast majority of other cell using a precisely targeted laser pulse. Instantly lysed cells release their contents into the medium, including mRNA, providing a method to analyze mRNA content in selected cells. Using a high throughput plate cytometer equipped with lasers for in situ cell manipulation (LEAP Cell Processing and Purification Workstation), we have analyzed RNA from individual tumor cells in a model of CTCs consisting of carcinoma cells spiked into human PBMCs. A 5-gene panel was measured by RT-PCR from single tumor cells and was detected in 94% of the cases with 0% false positives. RNA analysis of single tumor cells via laser-mediated lysis represents a new principle for the analysis of rare cells, as the specificity of analysis is dependent on imaging and laser targeting only and free of any limitations imposed by an antibody- or size-dependent purification strategy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 1162.