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Research

The Center for Bioimaging (CeBI) serves as a central research infrastructure for advanced light microscopy, spectral flow cytometry, and image analysis within the Department of Chemistry and Chemical Biology at TU Dortmund University. In addition to providing access to state-of-the-art imaging and analysis technologies, CeBi offers expect support for investigations of cell biological processes. 

Rapid perturbation and real-time activity measurements in living cells

To uncover mechanisms, how cellular structures are organized in space and time, methods are required that enable direct monitoring and acute perturbation of key regulators. To reach this goal, novel generic approaches were developed to simultaneously analyze and modulate biochemical reactions inside living cells.

In particular, to induce rapid and prolonged perturbations of protein activities in the plasma membrane, methods were developed based on chemically-induced dimerization and photochemically-induced targeting to immobilized artificial receptors to directly “paint” stable network perturbations in living cells. (Video 1) [1]

Demo-Video © Leif Dehmelt​/​TU Dortmund
Video 1: Rapid and prolonged perturbation of intracellular signal networks via "Molecular Activity Painting". A pattern of the signal protein GEF-H1 in the shape of the letter “N” was "painted" at the plasma membrane via uncaging with a focussed beam of UV light (left panel). Plasma membrane localization of GEF-H1 induced the new formation of dynamic, myosin-based contractile structures (middle panel). Right panel: combined channels.

These perturbations can be combined with activity measurements, to uncover perturbation response relationships in signal networks.

Refenrence

[1] Chen X, Venkatachalapathy M, Kamps D, Weigel S, Kumar R, Orlich M, Garrecht R, Hirtz M, Niemeyer CM, Wu YW, Dehmelt L. (2017).
Molecular-Activity Painting": Switch-like, Light-Controlled Perturbations inside Living Cells.
Angew Chem Int Ed Engl 21(14):5311-6.