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US 11,906,721 B2Granted
Method and device for manipulating a beam path in a microscope, method for recording image stacks in a microscope
AssigneeLeica Microsystems CMS GmbH
InventorFlorian Fahrbach
Life of this patent
Estimating the term…
Abstract
A method for manipulating at least one beam path in a microscope includes ascertaining a refractive index of a sample arranged in a sample volume and/or of an optical medium arranged in the sample volume. At least one microscope parameter is set in dependence on the ascertained refractive index for manipulating the beam path.
Claims24
A method for manipulating at least one beam path in a microscope, the method comprising:
- determining a refractive index of a sample arranged in a sample volume and/or of an optical medium arranged in the sample volume;
- and
- setting at least one microscope parameter based on the determined refractive index for manipulating the beam path,
- wherein determining the refractive index of the sample and/or of the optical medium arranged in the sample volume comprises:
- (I) oblique irradiation of the sample volume with measurement light with respect to the optical axis using an optical arrangement;
- (II) reflecting the irradiating light at a reflective element provided in the sample volume at a first position;
- (III) imaging the reflected light onto a spatially resolving detector;
- (IV) evaluating the signal detected by the detector with respect to a size and/or offset of the reflected light on the spatially resolving detector;
- (V) displacing the reflective element along the optical axis to a second position and performing the method steps (III) and (IV);
- and
- (VI) determining the refractive index based on the size and/or the offset of the reflected light for the first position and the second position of the reflective element.
8 more elements
A method for recording image stacks in a microscope, the method comprising:
- manipulating at least one beam path by:
- determining a refractive index of a sample arranged in a sample volume and/or of an optical medium arranged in the sample volume, and setting at least one microscope parameter based on the determined refractive index for manipulating the beam path;
- and
- capturing a change in a position of the sample with respect to an optical arrangement and/or capturing changes in an excitation wavelength before the at least one microscope parameter is set,
- wherein determining the refractive index of the sample and/or of the optical medium arranged in the sample volume comprises:
- (I) oblique irradiation of the sample volume with measurement light with respect to the optical axis using the optical arrangement;
- (II) reflecting the irradiating light at a reflective element provided in the sample volume at a first position;
- (III) imaging the reflected light onto a spatially resolving detector;
- (IV) evaluating the signal detected by the detector with respect to a size and/or offset of the reflected light on the spatially resolving detector;
- (V) displacing the reflective element along the optical axis to a second position and performing the method steps (III) and (IV);
- and
- (VI) determining the refractive index based on the size and/or the offset of the reflected light for the first position and the second position of the reflective element.
9 more elements
An apparatus for manipulating at least one beam path in a microscope, the apparatus comprising:
- a refractive-index determination module for determining the refractive index of a sample and/or the refractive index of an optical medium arranged in a sample volume;
- and
- at least one beam path manipulator for setting at least one microscope parameter based on the determined refractive index for manipulation of the at least one beam path,
- wherein determining the refractive index of the sample and/or of the optical medium arranged in the sample volume comprises:
- (I) oblique irradiation of the sample volume with measurement light with respect to the optical axis using an optical arrangement;
- (II) reflecting the irradiating light at a reflective element provided in the sample volume at a first position;
- (III) imaging the reflected light onto a spatially resolving detector;
- (IV) evaluating the signal detected by the detector with respect to a size and/or offset of the reflected light on the spatially resolving detector;
- (V) displacing the reflective element along the optical axis to a second position and performing the method steps (III) and (IV);
- and
- (VI) determining the refractive index based on the size and/or the offset of the reflected light for the first position and the second position of the reflective element.
8 more elements
A method for recording image stacks in a microscope, the method comprising:
- displacing a focal plane of a first optical arrangement by a preset scanning distance;
- manipulating at least one beam path of the first optical arrangement for correcting imaging aberrations of the first optical arrangement by:
- determining a refractive index of a sample arranged in a sample volume and/or of an optical medium arranged in the sample volume, and setting at least one microscope parameter based on the determined refractive index for manipulating the beam path;
- and
- displacing or adjusting the focal plane of a second optical arrangement by an adjustment distance that is based on the determined refractive index,
- wherein determining the refractive index of the sample and/or of the optical medium arranged in the sample volume comprises:
- (I) oblique irradiation of the sample volume with measurement light with respect to the optical axis using the first optical arrangement;
- (II) reflecting the irradiating light at a reflective element provided in the sample volume at a first position;
- (III) imaging the reflected light onto a spatially resolving detector;
- (IV) evaluating the signal detected by the detector with respect to a size and/or offset of the reflected light on the spatially resolving detector;
- (V) displacing the reflective element along the optical axis to a second position and performing the method steps (III) and (IV);
- and
- (VI) determining the refractive index based on the size and/or the offset of the reflected light for the first position and the second position of the reflective element.
10 more elements
A method for manipulating at least one beam path in a microscope, the method comprising:
- determining a refractive index of a sample arranged in a sample volume and/or of an optical medium arranged in the sample volume;
- and
- setting at least one microscope parameter based on the determined refractive index for manipulating the beam path,
- wherein determining of the refractive index of the sample and/or of the optical medium arranged in the sample volume comprises the following method steps:
- (i) focusing measurement light into the sample volume by an optical arrangement, wherein the measurement light is transmitted on a sample side of the optical arrangement by the optical medium and a further optical medium;
- (ii) detecting the measurement light that is reflected by a reflective element and transmitted by a further optical arrangement or by the optical arrangement, using a detector arrangement or using a spatially resolving detector;
- (iii) determining a working distance between the optical arrangement and the reflective element based on the measurement light detected by the detector, wherein, for the working distance, the focus of the measurement light lies on the reflective element;
- (iv) varying at least one of the following parameters:
- (iv.1) distance between the optical arrangement and the sample medium;
- (iv.2) distance between the reflector and the further optical medium;
- (iv.3) divergence of the measurement light,
- wherein the varying results in a fixed distance change of a focus position of the measurement light;
- (v) determining a further working distance according to method steps (i) to (iii);
- (vi) determining a working distance change between the working distance and the further working distance;
- and
- (vii) determining the refractive index based on the distance change and the working distance change.
13 more elements
Dependencies are inferred from the claim text.