CORDIS provides links to public deliverables and publications of HORIZON projects.
Links to deliverables and publications from FP7 projects, as well as links to some specific result types such as dataset and software, are dynamically retrieved from OpenAIRE .
Deliverables
Simultaneous multi-parameter Picosecond Fluorescence Lifetime Imaging Microscopy (PFLIM) has been developed, based on the novel concept of Time- and Space-Correlated Single Photon Counting (TSCSPC), which was made possible by the recent introduction of time- and space-sensitive microchannel plate (MCP) photomultipliers with delay-line (DL) and quadrant anodes (QA). The QA-detector has a space resolution of 100 micron, resulting in 200x200-space channelsat < 10 ps time resolution and an unprecedented dynamic range of 107. Simultaneous acquisition of laser-correlation time, absolute arrival time, xy-coordinates, emission wavelength, and polarization of each individual photon is possible. From the stored list-mode data, any combination of parameters can be obtained resulting, e.g., in sub-second frame-rate movies of mobile fluorescent particles including picosecond dynamics along the trajectory of the moving object. This capacity led to the development of Vehicle-Based Micro-Spectroscopy (VBMS), allowing it to follow the movement and interaction of a suitably labelled virus in living cells.
Photobleaching and photodynamic reactions do not distort the obtained data, since these effects can be separated in time, due to the movie-style acquisition. Biophysical information in living cells could be obtained under non-invasive conditions, such as ultra-low excitation light and ultra-low labeling, due to ultra-sensitive detection: (i) By observing the fluorescence dynamics at parallel and perpendicular orientation, fluorescence anisotropy decays of intercalated ethidium were obtained, revealing the chromatindynamics in the nucleus of living cells (Fig.1), (ii) DASPMI-labelled living cells allowed to obtain 3-exponential fluorescence dynamics (100 ps - 3 ns) of individual mitochondria (Fig.2), yielding new insights in the structural organization of the inner membrane, (iii) Fused GFP-proteins were expressed in living cells, whose fluorescence anisotropy dynamics revealed its 3D environment (Fig.3), and (iv) Understanding of photosynthesis in living plant leaves was advanced by comparing the 3-exponential fluorescence decays of individual chloroplasts in individual cells (Fig.4), at varying environmental conditions.
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