Prof. Ulrich Kubitscheck
The Biophysical Chemistry Workgroup, Rheinische Friedrich-Wilhelms Universität Bonn, Germany
The lab of Prof. Kubitscheck performs a wide range of imaging across different applications including single-molecule tracking and localization, where they examine the export of ribosomal particles through nuclear pore complexes by a number of methods. These methods include indirectly HaloTag labeling proteins that become part of pre-ribosomal subunits and imaging the DEAD-box helicase protein DBP5 that has a role in regulating the export of mRNA and pre-ribosomal particles. DBP5 is known to remove transport factors from mRNA in nuclear pore complexes, but further research is needed in order to uncover the molecular mechanisms and kinetics behind this export process.
As outlined by Prof. Kubitscheck, “The DBP5 protein diffuses around in the cell, attaches to the nuclear pore complexes and dissociates, these processes occur extremely rapidly and the challenge is to catch that in real-time.” This imaging application requires a combination of both speed and sensitivity, in order to capture the complex kinetics of the DBP5 protein while retaining a good signal to noise ratio to observe the samples and perform quantitative analysis. Furthermore, the nuclear pore complexes need to be visualized with high precision.
In addition, imaging was limited to a small field of view (FOV) due to the previous EMCCD solution requiring greater magnification.
“With the Prime BSI the signal to noise ratio is much better, and simultaneously the field of view is huge compared to the previous EMCCD.”