摘要:
A method for the three-dimensional imaging of a sample in which image information from different depth planes of the sample is stored in a spatially resolved manner, and the three-dimensional image of the sample is subsequently reconstructed from this stored image information is provided. A reference structure is applied to the illumination light, at least one fluorescing reference object is positioned next to or in the sample, images of the reference structure of the illumination light, of the reference object are recorded from at least one detection direction and evaluated. The light sheet is brought into an optimal position based on the results and image information of the reference object and of the sample from a plurality of detection directions is stored. Transformation operators are obtained on the basis of the stored image information and the reconstruction of the three-dimensional image of the is based on these transformation operators.
摘要:
The invention relates to a sample holding system for a microscope, comprising a sample chamber which has an upper opening and is filled with an immersion liquid, and in which a sample embedded in a transparent embedding compound is placed in a holder. The sample holding system also comprises means for the translatory movement of the sample in relation to a detection objective of the microscope, and means for rotating the sample about an essentially vertical rotational axis in a plane forming an angle different from zero with the optical axis of the detection objective. The means for rotating the sample in such a sample holding system comprise a rotational drive provided with a magnetic coupling or a belt drive and/or toothed wheel rotational drive arranged above the sample chamber, said drives transmitting the rotational movement to the holder, or the rotational movement is generated directly by the movement of the entire sample chamber.
摘要:
A sample holding system for a microscope, including a sample chamber which has an upper opening and is filled with an immersion liquid, and in which a sample embedded in a transparent embedding compound is placed in a holder. Translatory movement of the sample in relation to a detection objective of the microscope, and rotating the sample about an essentially vertical rotational axis in a plane forming an angle different from zero with the optical axis of the detection objective is included. Rotating the sample includes a rotational drive provided with a magnetic coupling or a belt drive and/or toothed wheel rotational drive arranged above the sample chamber.
摘要:
The invention is directed to method for positioning and aligning a preferably biological sample in the detection area of the objective of a microscope arrangement. According to the invention, the method mentioned above has the following method steps: a sample is introduced into a transparent medium, preferably agarose gel, which is initially liquid; the medium is changed from the liquid state to the solid state, wherein the sample is fixated within the medium, but the transparency of the medium is retained; the solidified medium is positioned in the microscope arrangement in such a way that the sample enclosed therein is situated in the detection area of the objective. Further, a device is proposed for positioning and aligning a preferably biological sample in the detection area of the objective of a microscope arrangement.
摘要:
The invention relates to methods for positioning at least one preferably biological specimen in the specimen space of a microscope arrangement, and to devices for carrying out these methods. Methods and devices are proposed, wherein the specimen's orientation relative to a detection objective's optical axis can be repeatedly changed and, in doing so, the specimen is held so that a substantially unobstructed view of the specimen is ensured from every detection direction. In different constructional variants, the specimen is held at a supporting device by adhesive forces or by a flowing medium, the specimen is held at a capillary opening by capillary action, or at least one specimen is embedded in a body of transparent gel, and the gel body is fixed in the specimen space by means of a rotatable holding device, and the detection direction is changed by rotating the holding device by a given angle of rotation.
摘要:
The invention is directed to an optical arrangement for photomanipulation of a sample comprising a sample holder for receiving the sample, an illumination device comprising an illumination light source and an illumination beam path for illuminating the sample with a light sheet. It further comprises a detection device for detecting light that is radiated from the sample and imaging optics which image the sample on the detection device by means of an imaging objective in an imaging beam path, wherein the light sheet is substantially planar in the focus of the imaging objective, and wherein the imaging objective has an optical axis which intersects the plane of the light sheet at an angle different from zero. Further, the arrangement also has means for photomanipulation of the sample.
摘要:
The invention relates to methods for positioning at least one preferably biological specimen in the specimen space of a microscope arrangement, and to devices for carrying out these methods. Methods and devices are proposed, wherein the specimen's orientation relative to a detection objective's optical axis can be repeatedly changed and, in doing so, the specimen is held so that a substantially unobstructed view of the specimen is ensured from every detection direction. In different constructional variants, the specimen is held at a supporting device by adhesive forces or by a flowing medium, the specimen is held at a capillary opening by capillary action, or at least one specimen is embedded in a body of transparent gel, and the gel body is fixed in the specimen space by means of a rotatable holding device, and the detection direction is changed by rotating the holding device by a given angle of rotation.
摘要:
The invention is directed to an optical arrangement with a light source for emitting a light bundle and with optical elements for transforming this light bundle into the shape of a light sheet, particularly suitable for illuminating individual planes of a three-dimensional specimen in selective plane illumination microscopy (SPIM). According to the invention, means are provided for varying the cross section of the light sheet, for varying the length of the light sheet and/or for influencing the direction in which individual beam components extending within the light sheet are directed to the specimen substance. This makes it possible to adapt the geometry of the light sheet to the illumination requirements for observing one and the same specimen plane with a plurality of different objectives and, if required, to reduce shadows occurring within the observed specimen plane as a result of the illumination.
摘要:
The invention is directed to a sample holder for a microscope. The sample holder comprises a sample chamber which is filled with an immersion liquid and in which a sample is located. The sample chamber has an upper opening. It further comprises means for translating the sample relative to a detection objective of the microscope, and means for rotating the sample around an axis of rotation extending in a substantially horizontal plane which encloses an angle other than zero degrees with the optical axis of the detection objective. In a sample holder of this kind, the sample is embedded in a transparent embedding medium having at least partially a greater solidity than the immersion liquid. Further, the sample chamber has means for horizontally supporting the embedded sample against the effect of gravity.
摘要:
Apparatus and method for high spatial resolution imaging of a sample's structure, including a diffraction-limited resolution volume with a plurality of dye molecules which can be switched between different states and have a distribution density which is greater than the inverse of the diffraction-limited resolution volume, where at least one state is fluorescing, the fluorescence being collected by an objective lens and imaged on a spatially resolving detector by an optical system. At least one light source provided for emitting a switching radiation and for emitting an excitation radiation. At least one of the light sources is arranged to radiate through the sample, and a switching and/or fluorescence excitation of the dye molecules is carried out. The switching is a photoactivation or a photodeactivation of the dye molecules. A focusing arrangement is provided for switching and/or for excitation to generate a line-like illumination region extending in a direction of illumination.