Arrangement for light sheet microscopy

Patent metadata

Field	Value
Publication number	US-10048482-B2
Application number	US-201415036510-A
Country	US
Kind code	B2
Filing date	Nov 13, 2014
Priority date	Nov 15, 2013
Publication date	Aug 14, 2018
Grant date	Aug 14, 2018

Abstract

An arrangement for light sheet microscopy including illumination optics with an illumination objective for illuminating a sample, located in a medium on a sample carrier aligned with respect to a plane reference surface, with a light sheet. The arrangement further includes detection optics with a detection objective. The arrangement also includes a separating layer system with at least one layer separating the medium from the illumination and detection objectives. The separating layer system is aligned with a base surface parallel to the reference surface and contacts the medium by this base surface. At least one detection corrective lens, for reducing those aberrations occurring as a result of light to be detected passing obliquely through interfaces of the separating layer system, is configured as free-form lens and is arranged between the detection objective and the separating layer system. Alternatively, it also forms the front lens of the detection objective.

First claim

The invention claimed is:

1. An arrangement for light sheet microscopy comprising:
illumination optics comprising:
an illumination objective configured to illuminate a sample with a light sheet via an illumination beam path, when the sample is located in a medium on a sample carrier that is aligned with respect to a plane reference surface;
wherein an optical axis of the illumination objective and the light sheet lies in a plane which forms an illumination angle not equal to zero with normal lines of the plane reference surface;

detection optics comprising:
a detection objective in a detection beam path;
wherein and optical axis of the detection objective forms a detection angle not equal to zero with the normal lines of the reference surface;

a separating layer system comprising:
at least one layer made of a predetermined material with a predetermined thickness which separates the medium from the illumination objective and the detection objective;
wherein the separating layer system is configured to be aligned with and contact the medium by a base surface parallel to the reference surface at least in a region that is accessible by the illumination objective and the detection objective for illuminating and detecting; and

a detection corrective lens system comprising:
at least one detection corrective lens configured to reduce aberrations that occur as a result of light to be detected passing obliquely through interfaces of the separating layer system;
wherein the at least one detection corrective lens is configured as a free-form lens, and is arranged between the detection objective and the separating layer system or forms a front lens of the detection objective.

2. The arrangement according to claim 1 ;
wherein the detection objective has a detection focusing device for internal focusing with simultaneous correction of spherical aberrations occurring during change of focus.

3. The arrangement according to claim 2 ;
wherein the detection focusing device includes at least a first adaptive optical detection focusing element that is arranged in, or can be inserted into, the beam path, configured to internally focus and simultaneously correct spherical aberrations occurring during a change of focus.

4. The arrangement according to claim 3 ;
wherein the detection objective comprises:
a first detection lens group arranged downstream of the detection corrective lens configured to image an object-side image field to infinity and
a second detection lens group; and

wherein the at least one adaptive optical detection focusing element is arranged between the first detection lens group and the second detection lens group.

5. The arrangement according to claim 1 , further comprising:
an illumination corrective lens system comprising at least one illumination corrective lens configured to reduce aberrations occurring as a result of illumination light passing obliquely through interfaces of the separating layer system;
wherein the illumination corrective lens is designed as a free-form lens, and is arranged between illumination objective and separating layer system or forms the front lens of the illumination objective.

6. The arrangement according to claim 5 ;
wherein the illumination objective comprises an illumination focusing device configured to internally focus and simultaneously correct spherical aberrations occurring during a change of focus.

7. The arrangement according to claim 6 ;
wherein the illumination focusing device includes at least one adaptive optical illumination focusing element that is arranged in, or is configured to be inserted into, the beam path for internal focusing and simultaneous correction of spherical aberrations occurring during a change of focus.

8. The arrangement according to claim 6 ;
wherein the illumination objective comprises:
a first illumination lens group, downstream of the illumination corrective lens, which is configured to image an object-side image field to infinity; and
a second illumination lens group; and

wherein the at least one adaptive optical illumination focusing element is arranged between the first illumination lens group and the second illumination lens group.

9. The arrangement according to claim 3 ;
wherein the at least one adaptive optical detection focusing element and the at least one illumination focusing element are formed as wavefront manipulators.

10. The arrangement according to claim 4 ;
wherein the second detection lens group is constructed in an afocal manner;
wherein a third detection lens group is formed as a tube lens group and is formed as a tube lens group and is arranged downstream of the second detection lens group; and
wherein the detection objective is configured to generate a real intermediate image.

11. The arrangement according to claim 4 ;
wherein the first detection lens group with the detection corrective lens and the at least one layer of the separating layer system, and the first illumination lens group with the illumination corrective lens and the at least one layer of the separating layer system, respectively, forms an optical system which approximately meets the following condition:
h 1 =f FG ·sin σ 0 =n 0 ·f FG ′·sin σ 0 ;

wherein:
h 1 is the incident height of the opening beam at the wavefront manipulator;
σ 0 is the angle of inclination of the edge beam relative to the optical axis;
f FG =−n 0 *f FG is the front object-side focal length of the front lens group; and
n 0 is the refractive index of an immersion medium between object and front lens.

12. The arrangement according to claim 11 ;
wherein the first detection lens group and/or the first illumination lens group comprise(s) an aspherical lens.

13. The arrangement according to claim 2 ;
wherein a numerical aperture NA of the detection objective is at least 0.8.

14. The arrangement according to claim 6 ;
wherein the numerical aperture NA of the illumination optics is at most 0.6.

15. The arrangement according to claim 9 ;
wherein each of the wavefront manipulators comprises two free-form surfaces that are movable opposite one another lateral to the optical axis.

16. The arrangement according to claim 15 ;
wherein an immersion medium is introduced in each instance between the free-form surfaces of the wavefront manipulators which are movable opposite one another.

17. The arrangement according to claim 15 ;
wherein the optical elements, at which the free-form surfaces are formed, are fabricated from a material with anomalous partial dispersion.

18. The arrangement according to claim 16 ;
wherein the immersion medium has a dispersion curve which deviates from the normal line.

Assignees

• Zeiss Carl Microscopy Gmbh

Inventors

• Pretorius Marco

Classifications

• G02B21/367

  providing an output produced by processing a plurality of individual source images, e.g. image tiling, montage, composite images, depth sectioning, image comparison · CPC title

• G02B21/08 Primary

  Condensers · CPC title

• G02B21/0076

  arrangements using fluorescence or luminescence · CPC title

• G02B21/0052

  Optical details of the image generation · CPC title

• G02B27/0025 Primary

  for optical correction, e.g. distorsion, aberration · CPC title