SP5

Technology development for TMM classification

Confocal screen of tissue micro arrays (TMAs): tissue sections are arranged in a 2D matrix pattern on a slide and imaged confocally in 3D.

Within subproject SP5 we deal with the following six tasks: (i) Development of protocols for visualization of cell biologically relevant features of Alternative Lenghtening of Telomeres (ALT) on tissue microarrays (TMAs). (ii) Evaluating additional Telomere Maintenance Mechanism (TMM) features such as C-circles that are characteristic for ALT cells to improve the model of SP3. (iii) Optimizing the TMM classification so that it can be applied robustly to a broad range of samples including various types of TMAs. (iv) Increasing the throughput of the automated image aquisition, which is currently a limiting factor for aquiring three-color image stacks of large numbers of cells as needed for good statistics to resolve all changes in telomere features. (v) Testing different methods for developing an optimized and cost efficient TMM analysis scheme that can be applied in a clinical setting, e.g. flow cytometry (FLOW FISH) and PCR for telomere length analysis as well as TERRA level measurement by RT-PCR. (vi) Developing novel strategies to inhibit the TMM for therapeutic intervention.

Activities of the ViroQuant-CellNetworks RNAi screening Facility.

Keywords: High-throughput screening; High-content-screening; siRNA