Imaging Cycler® technology

The following is a brief description of the know-how which has been established in ToposNomos Ltd. based on more than 20 years of research. ToposNomos Ltd. (TNL) provides the Imaging Cycler® technology as a hardware and software system for customers and project partners through its human toponome project (see HUTO project). Moreover, as a consultant TNL provides all the required know-how for customers to establish their own Toponome projects, both technologically and biologically in a cost and time effective manner. Imaging Cycler® microscopes are fully automated microscopic imaging and analysis systems allowing the researcher to colocalize a quasi random number of different molecular classes in one biological sample (cell or tissue section) by using large tag libraries. This ability is based upon repetitive cycles (more than 100) of fluorescence protein tagging and imaging in situ. Thereby ever Imaging Cycler® or large, interacting Imaging Cycler® clusters collects a large number of molecular distribution patterns in one sample by breaking the spectral limitations of traditional fluorescence microscopy and allowing to (functionally and multimolecularly) resolve structures known to be present at approx. 40 nm distance or less (Schubert W et al N Biotechnol, 2012). By aligning the signals, it detects the higher order combinatorial molecular features of protein systems in cells for the first time. It provides insight into the organized proteome (the toponome), in which the proteins are coordinately arranged as networks in time and space to generate function. Imaging Cycler® is applicable as high-throughput systems unraveling lead proteins controlling the hierarchical topology and function of protein networks in cells and tissues. This not only allows the researcher to uncover which and when proteins are co-sorted in cells and which are excluded, it also unravels key target proteins (pathological lead proteins) generating and controlling pathological protein networks in disease. These networks can be directly imaged as in situ Toponome fingerprints of diseases (combinatorial biomarkers, or TIS™ codes). Proteins occurring as lead proteins under disease conditions are key target candidates for drug developments. Moreover the technology underlying Imaging Cycler® is the first comprehensive basis for a microscopy with functional resolution. It provides insight into the cell at unprecedented detail simultaneously in all its organelles and subcellular structures. (Schubert W et al N Biotechnol, 2012)

Imaging Cycler® is the company´s proprietary technology. Patents cover any modification of cyclical fluorescence imaging (US 6,150,173; JP 3739528; EP 0180428; DE 19709348).

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History of the technology

The basic method for the dimension- and parameter- unlimited imaging of high-dimensional "molecular combination patterns" at subcellular resolution, essential for the quantitative analysis of protein network structures in cells, was developed by W. Schubert in 1996 (US 6,150,173; JP 3739528; EP 0180428; DE 19709348). The corresponding cyclical labelling procedure was then further developed technically by the Molecular Pattern Recognition Research (MPRR) group, headed by W. Schubert, at the University of Magdeburg, Institute of Medical Neurobiology, Germany. The technology was topic of research projects aiming at toponome biology and function. They were funded by the German BMBF, the DFG, the Land Saxony-Anhalt, the Klaus Tschira foundation and FP7 EU project. MPRR Key people in toponomics: Ch. Rethfeld, M. Bode, M. Friedenberger, A. Krusche, Reyk Hillert, A. Gieseler in close cooperation with W. Schubert.

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Wakefield, Munich, Magdeburg