Optical system for a light sheet microscope
An optical system for a light-sheet microscope comprises transporting optics configured to project, into a sample, a light sheet for illuminating a sample plane positioned obliquely to an optical axis of the transporting optics and to project the illuminated sample plane into an intermediate image space. The transporting optics comprises an interchanging system that includes a first light-deflection element and a second light-deflection element. The interchanging system is configured to switch an illumination direction along which the light sheet illuminates the sample by alternately introducing the first light-deflection element and the second light-deflection element into a beam path of the transporting optics. The first light-deflection element causes a partial image inversion in only one direction. The second light-deflection element causes a complete image inversion in two directions.
1. An optical system for a light-sheet microscope, the optical system comprising:
transporting optics configured to project, into a sample, a light sheet for illuminating a sample plane positioned obliquely to an optical axis of the transporting optics and to project the illuminated sample plane into an intermediate image space,
wherein the transporting optics comprises an interchanging system that includes a first light-deflection element and a second light-deflection element, the interchanging system being configured to switch an illumination direction along which the light sheet illuminates the sample by alternately introducing the first light-deflection element and the second light-deflection element into a beam path of the transporting optics, so that the light sheet illuminates the sample from a first direction when only the first light-deflection element is in the beam path, and the light sheet illuminates the sample from a second direction when only the second light-deflection element is in the beam path, the second direction being different from the first direction, and
wherein the first light-deflection element causes a partial image inversion in only one direction and the second light-deflection element causes a complete image inversion in two directions.
2. The optical system according to claim 1 , wherein the transporting optics are configured to project into the sample the light sheet generated in the intermediate image space for illuminating the sample plane positioned obliquely to the optical axis of the transporting optics.
3. The optical system according to claim 1 , further comprising detection optics, wherein an optical axis of the detection optics is tilted with respect to the optical axis of the transporting optics about a tilt axis that extends through the intermediate image space.
4. The optical system according to claim 3 , further comprising a scanning device for successive illumination, by the light sheet, of a plurality of sample planes arranged successively in a scanning direction, wherein the detection optics) are configured in each case to generate a tomographic image from an image of a respective sample plane generated in the intermediate image space.
5. The optical system according to claim 4 , wherein the scanning device comprises a grid mirror configured to move the light sheet transversely to the optical axis of the transporting optics.
6. The optical system according to claim 4 , wherein the scanning device comprises a focusing apparatus for adjusting a sharpness plane of the transporting optics.
7. The optical system according to claim 4 , further comprising an image processing and control unit configured to introduce the first light-deflection element into the beam path of the transporting optics to generate a first image dataset which represents a first tomographic image stack compiled from the tomographic images, and to introduce the second light-deflection element into the beam path of the transporting optics to generate a second image dataset which represents a second tomographic image stack compiled from the tomographic images, and to process the first image dataset and the second image dataset to form a three-dimensional image of the sample.
8. The optical system according to claim 1 , wherein the transporting optics has a single object lens facing the sample, an optical axis of the object lens being parallel to a tilt axis that extends through the intermediate image space.
9. The optical system according to claim 1 , wherein each of the first light-deflection element and the second light-deflection element deflects the beam path of the transporting optics by 90°.
10. The optical system according to claim 1 , wherein the first light-deflection element comprises a deflecting mirror.
11. The optical system according to claim 1 , wherein the second light-deflection element is a roof prism.
12. The optical system according to claim 1 , wherein the second light-deflection element causes an image rotation through 90°.
13. A light-sheet microscope comprising the optical system according to claim 1 .
14. The light-sheet microscope according to claim 13 , wherein the light-sheet microscope is an inverted microscope.
15. A method for illuminating a sample in a light-sheet microscope, the method comprising:
projecting, using transporting optics, a light sheet into a sample to illuminate a sample plane positioned obliquely to an optical axis of the transporting optics, wherein the illuminated sample plane is projected into an intermediate image space,
switching an illumination direction along which the light sheet illuminates the sample by alternately introducing a first light-deflection element and a second light-deflection element into a beam path of the transporting optics, so that the light sheet illuminates the sample from a first direction when only the first light-deflection element is in the beam path, and the light sheet illuminates the sample from a second direction when only the second light-deflection element is in the beam path, the second direction being different from the first direction,
wherein the first light-deflection element causes a partial image inversion in only one direction and the second light-deflection element causes a complete image inversion in two directions.
16. The optical system according to claim 1 , wherein the first light-deflection element comprises a deflecting prism.
17. The optical system according to claim 1 , wherein the second light-deflection element comprises a roof mirror, or an Amici prism.
18. The optical system according to claim 1 , wherein the second light-deflection element comprises an Amici prism.
19. The optical system according to claim 1 , wherein the second light-deflection element causes an image rotation through 180°, or 270°.
20. The optical system according to claim 1 , wherein the second light-deflection element causes an image rotation through 270°.