Stabilization of focus in high resolution microscopy

Feb. 14, 2019

Stabilization of focus in high resolution microscopy

Technology description

Monitoring and correcting relative position without disturbing the use of the microscope objective.
Test beam directed onto a reflective surface connected to the sample/objective evaluated.
Necessary for ultra-high resolution microscopy with increasing spatial resolution

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.

Solutions

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.

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.

It depicts a microscope 1 including a microscope objective 2. The microscope objective 2 comprises a front lens 3 and a frame 4 holding the front lens 3 and having a metallic surface 5. In the operation of the microscope 1, a beam of excitation light 6 is focused into a sample 7 by means of the front lens 3. The focus of the beam of excitation light 6 is moved along a focal plane of the front lens 3 by means of a scanning device of the microscope 1 , which is not depicted here. The focal plane of the microscope objective 2 only remains fixed with regard to the sample 7, if relative positions of the sample 7 and of a microscope slide 8 and a cover slip 9 between which the sample 7 is fixed remain fixed with regard to the microscope objective 2.

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