Leica Microsystems Cms Gmbh

Andreas Hecker

Werner Knebel

Heinrich Ulrich

Kyra Moellmann

A microscope for conventional fluorescence microscopy (epi-fluorescence) and for total internal reflection microscopy is described. A first light source emits conventional fluorescent illumination light along a first illumination path and a second light source emitting evanescent illumination light along a second illumination path that differs from the first illumination path. An objective emits light onto an object to be viewed. A beam combiner directs the two lights into the objective while keeping their beam paths geometrically separated. The beam combiner comprises at least two spatially separated first zones for coupling in the conventional fluorescent illumination light and at least two spatially separated second zones for coupling in the evanescent illumination light. The first and second zones are adapted in their size and position to objective pupils of different objectives.

Microscope for conventional fluorescence microscopy and total internal reflection microscopy

A microscope for total internal reflection microscopy. The microscope includes at least one light source configured to provide an illumination light to an illumination beam path for an evanescent illumination of a specimen so as to reflect the illumination light at an interface to the specimen or a specimen cover so as to return reflection light into the illumination beam path, an objective through which the illumination light and detection light are directable, a detection device, and a coupling device. The coupling device includes a mirror disposed in the illumination beam path. The mirror has a reflecting surface and a hole, the hole being configured to pass the illumination light there through so as to couple the illumination light into the illumination beam path. The reflecting surface is configured to couple out at least a part of the reflection light and to direct the coupled-out reflection light to the detection devices so as to enable determination from a beam path of the coupled-out reflection light, at least one of a quantifiable parameter and a qualifiable parameter of at least one of the evanescent illumination and an evanescent field created in the specimen.

Microscope and method for total internal reflection-microscopy

A microscope with a light source that produces an illumination light beam for evanescently illuminating a sample includes an adjustment mechanism with which the polarization of the illumination light beam may be changed.

Illumination module for evanescent illumination and microscope

A method of ascertaining the orientation of molecules in a biological specimen by total internal reflection includes focusing illuminating light through an objective in different positions in a plane of a pupil of the objective so as to generate a plurality of respective differently oriented evanescent fields in the specimen. Respective different fluorescence intensities resulting from the differently oriented evanescent fields are correlated with respective different orientations of molecules in the specimen.

Method for ascertaining the orientation of molecules in biological specimens

A microscope with evanescent sample illumination and a method for testing samples are disclosed. A first evanescent field, which exhibits a first penetration depth in the sample, and a second evanescent field, which exhibits a second penetration depth in the sample that is greater than the first penetration depth, are produced. A detector is provided that detects the first detection light, which exits from the part of the sample illuminated with the first evanescent field, and which produces first detection light data therefrom, and the second detection light, which exits from the part of the sample illuminated with the second evanescent field, and which produces second detection light data therefrom. Furthermore, a processing module is provided for processing the first and second detection light data.

Method for analyzing a sample and microscope for evanescently illuminating the sample

The invention relates to a microscope for conventional fluorescence microscopy (Epi fluorescence) and for total internal reflection microscopy, including at least one light source () for the conventional fluorescence illumination and at least one light source () for the evanescent illumination, and including an objective (), wherein the illuminating light coming from the light sources () on different illumination paths () enters the objective () via a beam combiner () and from there is passed to the sample (), characterized in that the exit pupil of the objective () is imaged on the beam combiner () and the beam combiner is structured such that it guides the illuminating light used for the conventional fluorescence illumination and the illuminating light used for the evanescent illumination into the objective () along beam paths () which are geometrically separate from one another, and preferably run parallel or coaxially to one another.

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Asslar, Germany

Andreas Hecker