Microscope for imaging a sample
A microscope is disclosed for imaging a sample. The microscope includes an illumination objective, a further illumination objective, an imaging objective and a further imaging objective, a sample holder, holder support and a drive system arranged to displace the sample holder along three perpendicular axes.
1. A microscope comprising:
a sample holder configured to receive a sample and having a light transparent portion;
a first illumination objective arranged to eject a first illumination light beam along a first illumination path to illuminate the sample received by the sample holder through the light transparent portion of the sample holder such that light is ejected from the sample;
a second illumination objective arranged to eject a second illumination light beam along a second illumination path to illuminate the sample received by the sample holder through the light transparent portion of the sample holder such that light is ejected from the sample;
a first imaging objective arranged to receive detection light comprising at least a portion of the light ejected from the sample, wherein the detection light is propagated along a first detection path angled at about 90° to the first and second illumination paths and wherein the first imaging objective has a first imaging focal plane;
a second imaging objective arranged to receive detection light comprising at least a portion of the light ejected from the sample, wherein the detection light is propagated along a second detection path angled at about 90° to the first and second illumination paths and wherein the second imaging objective has a second imaging focal plane; and
a holder support arranged to support the sample holder, wherein, relative to a direction of gravity, the first and second illumination objectives are arranged such that the first and second illumination path are located below a bottom of the sample holder,
wherein the first and second illumination paths are not parallel, and
wherein the holder support has a drive system arranged to displace the sample holder along two perpendicular axes relative to the first and second imaging objectives.
2. The microscope of claim 1 , wherein the first imaging focal plane and the second imaging focal plane are essentially vertically oriented along the direction of gravity.
3. The microscope of claim 1 , wherein the first imaging focal plane and second imaging focal plane overlap, and wherein the first illumination path and the second illumination path lie in the first imaging focal plane and the second imaging focal plane.
4. The microscope of claim 1 , wherein the first illumination path is angled between 10 degrees and 60 degrees to a horizontal plane, and wherein the second illumination path is angled between 10 degrees and 60 degrees to the horizontal plane.
5. The microscope of claim 1 , wherein the drive system of the holder support is configured to displace the sample holder so that at least a part of the sample holder is located directly above the first illumination objective and/or second illumination objective along the direction of gravity.
6. The microscope of claim 1 , wherein the light transparent portion of the sample holder contains the bottom of the sample holder and side walls of the sample holder, wherein the emitted light passes through the side walls of the sample holder and wherein the sample holder has an open top end.
7. The microscope of claim 1 , wherein the light transparent portion of the sample holder has a length along the X axis of at least two centimeters, and wherein the drive system of the holder support 1 S configured to displace the sample holder at least two centimeters along the X axis.
8. The microscope of claim 1 , wherein the light transparent portion of the sample holder contains at least one indentation extending into the direction of gravity.
9. The microscope of claim 1 , wherein the sample holder contains separation walls.
10. The microscope of claim 9 , wherein the light transparent portion of the sample holder contains at least one indentation extending into the direction of gravity, and wherein a bottom end of the indentation of the sample holder is at the same level or below a bottom end of the separation walls of the sample holder.
11. The microscope of claim 9 , wherein the separation walls are light transparent, and/or
wherein the light transparent portion of the sample holder is made of a material attached to an outside of the separation walls.
12. The microscope of claim 1 , wherein the sample holder contains at least two pockets opened on top and extending into the direction of gravity wherein the light transparent portion forms at least the bottom and sides of the pockets and wherein each pocket is arranged to hold a different sample.
13. The microscope of claim 1 , comprising a removably arranged platform, wherein the first illumination objective, the second illumination objective, the first imaging objective and the second imaging objective are mounted on the platform.
14. The microscope of claim 1 , comprising an electronically controllable element, wherein the first illumination objective is configured to eject the first illumination light beam with a first beam waist and the second illumination objective is configured to eject the second light beam with a second beam waist, and wherein the electronically controllable element is configured to translate the first beam waist along the first illumination path and the second beam waist along the second illumination path.
15. The microscope of claim 14 , wherein a first sheet of light is generated by scanning of the first light beam in a direction perpendicular to the first illumination path and a second sheet of light is generated by scanning of the second light beam in direction perpendicular to the second illumination path, and wherein the first beam waist is translated along the first illumination path and the second beam waist is translated along the second illumination path in synchronization with the scanning such that the first beam waist is scanned along an axis which is parallel to an axis of scanning of the second beam waist.
16. The microscope of claim 14 ,
configured to acquire a plurality of images of the same position of the sample including
an image of the sample illuminated with the first light beam having the first beam waist at a first position,
an image of the sample illuminated with the first light beam having the first beam waist translated along the first illumination path to a different second position,
an image of the sample illuminated with the second light beam having the second beam waist at a first position, and
an image of the sample illuminated with the second light beam having the second beam waist translated along the second illumination path to a different second position, and
preferably configured to reconstruct a combined image from the plurality of images by combining portions of the plurality of images illuminated by the waist of the light beams.
17. The microscope of claim 1 , comprising:
a first focusing system arranged to translate the first imaging focal plane of the first imaging objective along the first detection path, and/or
a second focusing system arranged to translate the second imaging focal plane of the second imaging objective along the second detection path.
18. The microscope of claim 17 ,
wherein the first focusing system is electronically controlled, the second focusing system is electronically controlled, or the first and second focusing systems are electronically controlled, or
wherein the first focusing system is realized using an electronically controlled deformable mirror, the second focusing system is realized using an electronically controlled deformable mirror, or the first and second focusing systems are realized using an electronically controlled deformable mirror, or
wherein the first focusing system is realized using an electronically controlled lens with tuneable focal length, the second focusing system is realized using an electronically controlled lens with tuneable focal length, or the first and second focusing systems are realized using an electronically controlled lens with tuneable focal lengths.
19. The microscope of claim 1 , comprising a light source arranged to illuminate the sample through the first imaging objective and to collect the transmitted light through the second imaging objective to generate a transmission image of the sample; and
preferably comprising a light source arranged to illuminate the sample through the second imaging objective and to collect the transmitted light through the first imaging objective to generate the transmission image of the sample.
20. The microscope of claim 1 , wherein the drive system of the holder support is arranged to displace the sample holder along three perpendicular axes relative to the first and second imaging objectives.