Development of modular spectral confocal fluorescence microscope with enhanced spatial resolution, using combined EM-CCD and SPAD detection, and offering special FCS-based point measurement modes
Grant Agency
Grant Agency of the Academy of Sciences of the Czech Republic
Topics
Biospectroscopy.
Year from
2009
Year to
2011
Abstract:
The task of the project is to develop a new modular highly sensitive spectral confocal fluorescence microscope with enhanced spatial resolution, using combined EM-CCD and SPAD detection, and offering two-focus, lifetime and newly suggested spectral FCS point measurement modes. The enhanced spatial resolution will be achieved by implementing Dynamic Saturation Optical Microscopy (DSOM), using saturation of fluorescence molecules into triplet or reversibly deactivated state. The goal is to reach 2-3 times better resolution than diffraction limit. For dynamic and interaction analysis the newest FCS methods including spectral FCS will be implemented. The spectral FCS will enable measuring dynamics of fluorescence molecules with overlapping emission spectra. The modularity of the microscope should enable latter incorporation of PALM and STORM. The microscope will be applied for studies of both model systems and living cells within the framework of existing projects and collaborations.
The task of the project is to develop a new modular highly sensitive spectral confocal fluorescence microscope with enhanced spatial resolution, using combined EM-CCD and SPAD detection, and offering two-focus, lifetime and newly suggested spectral FCS point measurement modes. The enhanced spatial resolution will be achieved by implementing Dynamic Saturation Optical Microscopy (DSOM), using saturation of fluorescence molecules into triplet or reversibly deactivated state. The goal is to reach 2-3 times better resolution than diffraction limit. For dynamic and interaction analysis the newest FCS methods including spectral FCS will be implemented. The spectral FCS will enable measuring dynamics of fluorescence molecules with overlapping emission spectra. The modularity of the microscope should enable latter incorporation of PALM and STORM. The microscope will be applied for studies of both model systems and living cells within the framework of existing projects and collaborations.
