Berlin 2005 – wissenschaftliches Programm

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AKB: Biologische Physik

AKB 50: Imaging and Microscopy

AKB 50.6: Vortrag

Montag, 7. März 2005, 17:15–17:30, TU H2013

Subdiffraction Fluorescence Imaging with photoswitchable fluorescent proteins — •M. Hofmann, C. Eggeling, S. Jakobs, and S.W. Hell — MPI für biophysikalische Chemie, Dep. NanoBiophotonics

The resolution of optical imaging in conventional far-field microscopy is limited by the diffraction of light. We present fluorescence imaging beyond this barrier by controlling the light driven transition between the dark and fluorescent state of a photoschwitchable protein. Fluorescence emission in the outer region of a diffraction-limited excitation spot is deactivated in a saturated manner, thereby reducing the effective fluorescence volume. Scanning images of protein stained structures exhibit an increased resolution. This can be described on the basis of a photophysical model and its underlying rate constants, which were determined from spectroscopic experiments of the fluorescence emission. The data shows that a reversible saturable optical fluorescence transition of a protein can be utilized to achieve optical imaging beyond the diffraction limit.

Hell, S. W., Nature Biotechnol. 21(11): 1347-1355 (2003)

M. Hofmann, C. Eggeling, S. Jakobs, S. W. Hell "Subdiffraction Imaging with the photoswitchable fluorescent protein asCP" (in preparation)