High-throughput gene expression screening in activated T-cells using FISH-Flow

Induction of gene expression and subsequent detection by FISH-Flow. c-fos expression is induced in activated cells, which are readily identified by FISH-Flow. GFP: negative control, GAPDH: positive control. Invention Summary: Researchers at Rutgers have developed an assay combining the analytical power of flow cytometry and fluorescence in situ hybridization (FISH-Flow) for high-throughput identification of activated T-cells. Gene expression at the mRNA level and expression of cell surface markers can be assayed simultaneously in activated T-cell populations using this technique. The use of nucleic acid probes provides exceptional target flexibility while maintaining exquisite sensitivity; as few as 10 mRNA molecules per cell can be detected by FISH-Flow. After being analyzed by flow cytometry, cell populations can be sorted for additional downstream analysis and multiparametric phenotyping. A semi-automated protocol has also been developed which allows for preparation and analysis of up to 40 samples simultaneously. Market Applications: Suitable for use in diagnostic and/or research applications requiring any of the following: Rapid identification of activated T cells and/or identification of low-abundance T cell populations, such as antigen-specific memory cells Multiparametric immunophenotyping Concurrent detection of mRNA targets and cell surface markers Advantages: Enhanced temporal resolution of dynamic gene expression events Eliminates the need for protein-secretion inhibitors when detecting secreted proteins such as cytokines Nucleic acid probes provide greater target flexibility (vs. antibody probes) Semiautomated protocol reduces cell loss, operator time, and inter operator variability Intellectual Property & Development Status: Patent pending. Available for licensing and/or research collaboration.