TY  - GEN
ID  - heidok9588
Y1  - 2008///
KW  - confocal microscopy 
KW  -  microscopy 
KW  -  fluorescence correlation spectroscopy 
KW  -  molecular mobility 
KW  -  line confocal
AV  - public
UR  - https://archiv.ub.uni-heidelberg.de/volltextserver/9588/
N2  - A spatial and temporal fluctuation microscope (STFM) was developed for fast imaging as well as the measurement and visualisation of mobilites and interactions of fluorescently labeled particles. The instrument extends the concept of a point confocal laser-scanning microscope to illumination and detection along a line. For the construction and application of the STFM, theoretical descriptions of the beam geometry, the point spread functions and the fluorescence auto- and cross-correlation fit functions were derived. The final system provids fast confocal imaging up to 30 images per second and allows for the spatially resolved detection of particle concentration fluctuations down to 15 ?s time resolution at hundreds of points in parallel. An optical resolution of 200 x 380 x 700 nm in x-, y- and z-direction was determined experimentally, and the spatially resolved fluctuations of fluorescent quantum dots were evaluated. Finally, the capabilities of the STFM in terms of fast imaging and mobility measurements were applied to studies of PML nuclear bodies and green fluorescent protein in living cells. Both the in vitro and the in vivo fluorescence correlation spectroscopy experiments conducted with the STFM were compared to single point measurements with a commercial point-confocal microscope and FCS setup. The results demonstrate the unique capabilities of the STFM for the characterisation of position-dependent diffusive translocations of macromolecules in the cell with very high time resolution.
TI  - Development and design of a spatially and temporally resolved fluorescence fluctuation microscope for the analysis of molecular mobilities and interactions
A1  - Heuvelman, Gerrit Lambertus
ER  -