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Long-Period Observation of Single (Bio)-Molecular Motors by Minimal-Invasive Fluorescence Lifetime Imaging Nanoscopy (FLIN)

Objective

The recently invented fluorescence lifetime imaging nanoscopy (FLIN) provides a groundbreaking tool for the study of single molecules (SM) and single molecular motors (SMM) as well as a broad array of phenomena in the NanoWorld. Previous limitations for SM M studies, resolution, short observation times, and photo-dynamic reactions, are now overcome by minimal-invasive picosecond FLIN. FLIN is the extension of the extremely successful fluorescence lifetime imaging microscopy (FLIM) into the nano-domain, with 10 to 100 nm space resolution. FLIN results from the combination of 4pi-microscopy with novel ultrasensitive, nonscanning imaging detectors, based on time- and space-correlated single photon counting (TSCSPC) that allows ultra-low excitation levels. This r esults in long-period (#gt; 1 hour), minimal-invasive observation of living cells and SM/SMM, without any cell damage or irreversible bleaching. Minimal-invasive FLIN (MI-FLIN) with global point spread function modelling allows observation of SMM movement at 1-nm accuracy and 10-nm resolution. Parallel to (i) MI-FLIN/FLIM implementation, the consortium will (ii) improve sensitivity, time- and space-resolution as well as throughput of the TSCSPC detectors, (iii) explore an array of novel applications provide d by MI-FLIM/FLIN, such as nanometer SMM-tracking, (iv) develop a super-background-free TIRF microscope to improve detectability of SM/SMM, and (v) examine the behaviour of four different types of SMM and their dependence on energy-input. Enhanced basic un derstanding of biological and artificial machines and motors will lead to improved model systems and proceed one day to the design of artificial systems, improving the interface of biological and non-biological worlds. Furthermore, biological SMM are invol ved in many disease states such as Alzheimers, Werner syndrome and infectious diseases. Our studies aim to improve understanding of how these motors operate and how they break down in disease.

Keywords

Call for proposal

FP6-2003-NMP-TI-3-MAIN
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Coordinator

EUROPHOTON GMBH, GESELLSCHAFT FUER OPTISCHE SENSORIK
EU contribution
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Address
Mozartstrasse 27
BERLIN
Germany

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Participants (9)