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Kurzfassung

Many light emitting entities routinely used, in cell biological settings show a dipole orientation. If the exact z position of the light emitting entity is unknown, using simple two-dimensional Gaussian fit algorithms to the light intensity distribution of a range of emitted light, may lead to calculation errors of the x- and y-position of several 10nm. The invention discloses a method for calculation of the exact spatial position and orientation of a light emitting in a sample.

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Hintergrund

Many light emitting entities routinely used, in cell biological settings show a dipole orientation. If the exact z position of the light emitting entity is unknown, using simple two-dimensional Gaussian fit algorithms to the light intensity distribution of a range of emitted light, may lead to calculation errors of the x- and y-position of several 10nm. The invention discloses a method for calculation of the exact spatial position and orientation of a light emitting in a sample.

The method has been verified experimentally as well as theoretically (see reference below).  

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Bilder & Videos

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Lösung

First the virtual x- and y-positions and of a light emitting entity are determined for several plans parallel to the focus plan. Thereafter the absolute and/or relative total or peak emitted light intensities in the images of the marker entity are accounted for in determining the z-position and spatial orientation of the marker (see figure). 

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Vorteile

• Fluorescence super-resolution
• Single molecule localization accuracy 

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Anwendungsbereiche

DKFZ is looking for a licensee for further development and commercialisation this technology for samples in high resolution microscopes particularly FPALM, PALM, STORM and PALMIRA.