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Localization Light Emitting Molecules of Unknown Orientation

Ref-Nr: TA-P-899


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.


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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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). 


Vorteile

  • Fluorescence super-resolution
  • Single molecule localization accuracy 

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. 


Deutsches Krebsforschungszentrum DKFZ

Dr. Frieder Kern
+49-6221-42-2952
f.kern@dkfz.de
www.dkfz.de
Adresse
Im Neuenheimer Feld 280
69120 Heidelberg



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