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.
