Meetings

19.12.2008: Kickoff-Meeting in Gräfelfing

05.11.2009: 1st Annual Meeting in Gräfelfing (TILL Photonics)

28.04.2011: 2nd Annual Meeting in Linz (Johannes-Kepler-University)

Results of the first project period (01.12.2008 - 31.05.2010)

Light and atomic force microscopy are complementary techniques for the study of live cells. With light microscopy, the interior of living cell is accessible for observation via fluorescence emission, and when light microscopy is combined with optical tweezers, molecular interactions can be observed even inside an intact cell. In contrast, AFM provides resolution on the cell surface topography at the nanometre scale and allows for studying mechanical interactions of membrane components. With SMW, all conceivable functions of these complementary technologies shall be integrated into a single and unified Single Molecule Workstation (SMW) concept. The microscopy hardware to be developed will be combined with a single, highly intuitive graphical user interface (GUI) integrating all functional modules of the system, exchanging and controlling measurement protocols in real time and in highly synchronised fashion. The application of this SMW instrument will enable major advances in life science research by delivering new insights into the molecular biology of living cells and complex biomolecular processes, which are yet inaccessible.

The main objective of the project is to create a flexible, modular and user-friendly workstation combining the three most important microscopic techniques ultra-sensitive inverted light microscopy (ILM), atomic force microscopy (AFM) for studying cell topography and molecular recognition imaging, optical tweezers (OT) for the detection of ultra-low forces and additional photo-thermal nano-spectroscopy (PTNS) to allow for the chemical characterisation of cellular material. These tasks are worked on by the SMEs and the industrial partner of the consortium. On a second level, the SMW instrument will be applied for the study of the correlation between structure and function of living cells, with exemplary applications in molecular immunology and basic cancer research as performed by the academic partners.

The project started on 1 December 2008. During the first 18 months, significant advances have been achieved. After the conceptual design of the mechanical, optical and electronic concept for the combination of AFM and ILM, a first prototype of a combi AFM-ILM instrument was built and its functionality was successfully tested. Furthermore, the requirements for integration of OT and PTNS into the SMW platform have been designed and evaluated in practice. In parallel, concepts for the combined software were developed and implemented into a functional version of a combined software and GUI for basic and expert protocols.
The academic partners have established appropriate test systems for the application of the SMW platform to study immunological process or detect cancer and have run a set of experiments with the first combi AFM-ILM prototype.

Thus, the SMW project has been successfully started in the first period. In the second period, it is planned to validate and continuously optimize the combi AFM-ILM hardware and also the software will be optimized correspondingly. Consequently, the project will progress further towards its objectives in the forthcoming second period.

Simultaneous epi-fluorescence microscopy and topography AFM imaging. Left panel: overlay of fluorescence and topography AFM images. The scan size was 100x100μm2 with the height scale ranging from 0 to 0.45μm for left panel and the scan size was 100x100μm2 with the height scale ranging from 0 to 0.95 μm for right panel of hSGLT1 expressing CHO cells. Right panel: overlay fluorescence and topography AFM images. The scan size was 100x100μm2 with the height scale ranging from 0 to 2.4μm for left panel and the scan size was 60x60μm2 with the height scale ranging from 0 to 0.5μm for right panel (in zoomed area) of rhodamine-phalloidin stained MyEnd cells (figure taken from Duman et al., 2010).