Laserscanning Mikroscope

• Subject Area: Biological Chemistry and Food Chemistry
• Term: Funded in 2020
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 442216850

A confocal high resolution microscope system is required for spatial separation of molecular structures and for the rapid detection of multiple fluorescence staining signals. It is planned that this microscope system will be used by members of the Biochemical Institute, the Institute of Pharmacy, Institute of Physiology, Institute of Anatomy and members of the collaborative research center 877 (SFB877) of the University Kiel. In the framework of the CRC877 the system will be part of the central microscopy service platform. For live cell imaging an inverse microscope system with a chamber is needed allowing temperature regulation and continous gas supply. The microscope system is planned for fixed cells, tissue samples and living cells. For the detection of dymanic processes of cell communication it is required to detect different fluorescence signals simultanously and with a sufficient speed of recording. A very high resolution and a rapid detection mode is necessary allowing a minumum of sample loss due to low laser intensities. For this purpose a high pixel dwell time to image even very weak signals with high sensitivity and low signal/noice ratio in the live cell modus has to be applied. The microscope should allow convocal imaging to detect multicolor fluorescence signals in a precise spatial and non overlapping mode. The optical resolution of the system should be able to process and image very tiny intracellular and extracellular structures, the composition of microenviroments, colocalisation and interaction of fluorecent proteins (FRET). This requires a sensitive detection methodology combined with mathematical applications to analyse signals and images (e.g. deconvolution) to optimize the signal/noice ratio and signal quality, respectively. To image multiple data points, multiple cells and tissue areas with sufficient detail and in a reproducible way over longer periods a motorized scanning table is needed. The wavelengths for the detection of the emission should be individually determined and called up again at any time. For the differentiation of signals of multilabelling the emission requires detailed spectral analysis. In addition moduls for interaction studies, FRAP, photo activation and the localisation of markers for physiological studies using sensitive fluorescent biosensors are required by different users. To monitor dynamic processes in a live cell modus a fast scanning system to detect in parallel different markers is required.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Laserscanning Mikroskop
• Instrumentation Group: 5090 Spezialmikroskope
• Applicant Institution: Christian-Albrechts-Universität zu Kiel
• Leader: Professor Dr. Paul Saftig