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Stabilization of focus in high resolution microscopy

Ref-Nr: TA-P-1260


Kurzfassung

The relative position of a microscope objective with regard to a sample is essential in imaging the sample using the microscope objective. The invention describes a method in which monitoring and correcting of a relative position of a microscope objective with regard to a sample is easily realized and does not disturb the primary use of the microscope objective. The procedure is carried out using a test beam directed onto a reflective surface connected to the sample and on the other hand using a test beam directed onto a reflective surface connected to the objective. Both results are registered, evaluated and used for correction.


Hintergrund

The relative position of a microscope objective with regard to a sample is essential in imaging the sample using the microscope objective. If relative position varies unnoticed, the plane of the sample imaged using the microscope objective will also vary unnoticed. As a consequence, a series of images which are intended show the same sample location at consecutive points in time, for example, in fact show the sample at different focal planes or laterally shifted. Further, with any laser scanning microscopy requiring a longer period of time for even imaging one plane of the sample once, a distorted image of the sample will be generated, if the relative position of the microscope objective with regard to the sample varies. With increasing spatial resolution achieved by ultrahigh resolution microscopy techniques like STED the requirements to be fulfilled in keeping a fixed relative position of the microscope objective with regard to the sample increases further. The increased spatial resolution requires an equally increased stability of the relative position of the microscope objective with regard to the sample.

The invention describes a method in which monitoring and correcting of a relative position of a microscope objective with regard to a sample is easily realized and does not disturb the primary use of the microscope objective. The procedure is carried out using a test beam directed onto a reflective surface connected to the sample and on the other hand using a test beam directed onto a reflective surface connected to the objective. Both results are registered, evaluated and used for correction.


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Lösung

  • Using a test beam directed onto a reflective surface connected to the sample.
  • Using a test beam directed onto a reflective surface connected to the objective.

 

 


Anwendungsbereiche

  • The invention can be used in high resolution microscopy such e.g. in a STED microscope.
  • A prototype has been successfully tested in experiments, see figure

 

 


Deutsches Krebsforschungszentrum DKFZ

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



Entwicklungsstand

Prototyp


Stichworte

Microscopy

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