Optomechanical module for converting a microscope to provide selective plane illumination microscopy

Patent metadata

Field	Value
Publication number	US-9316824-B2
Application number	US-201214003380-A
Country	US
Kind code	B2
Filing date	Mar 2, 2012
Priority date	Mar 4, 2011
Publication date	Apr 19, 2016
Grant date	Apr 19, 2016

Abstract

A module for engagement with a conventional microscope to provide selective plane illumination microscopy is disclosed. The module is coupled to the translational base of the microscope and defines a mounting having a mount body in which an excitation objective having a first longitudinal axis is engaged to one portion of the mount body and a detection objective having a second longitudinal axis is engaged to another portion of the mount body such that the first longitudinal axis is in perpendicular geometric relation with the second longitudinal axis.

First claim

What is claimed is:

1. A module for converting a microscope to provide selective plane illumination microcopy comprising:
a mount defining a mount body;
an excitation objective having a first longitudinal axis engaged to one portion of the mount body and a detection objective having a second longitudinal axis engaged to another portion of the mount body, wherein the excitation objective and the detection objective are engaged to the mount such that the first longitudinal axis intersects the second longitudinal axis in a perpendicular geometric relation, and
a light source for generating in alternating fashion a first plurality of light sheets through the excitation objective and then a second plurality of light sheets through the detection objective, and vice versa, for producing an excitation in a sample that generates an alternating sequence of a first fluorescent emission detected by the excitation objective and then a second fluorescent emission detected by the detection objective, and vice versa.

2. The module of claim 1 , wherein the mount body includes a first arm engaged to the excitation objective and a second arm engaged to the detection objective.

3. The module of claim 1 , wherein the light source forms a part of a selective plane illumination microscopy (SPIM) arrangement for generating the first plurality of light sheets and the second plurality of light sheets.

4. The module of claim 1 , wherein the mount is configured to be engaged to a microscope.

5. The module of claim 1 , wherein the microscope comprises:
a base configured to engage a sample stage for supporting and orienting the sample in an x-y direction; and
a translational stage configured to engage the stage for orienting the sample in the z-direction.

6. The module of claim 5 , wherein the microscope further comprises:
a conventional objective arrangement in communication with the base for detecting an illumination of the sample at an orientation different from the detection objective.

7. The module of claim 1 , wherein the mount has a first arm and a second arm having a perpendicular configuration in which the first arm and second arm are oriented perpendicularly to each other.

8. The module of claim 1 , further comprising:
a piezoelectric objective stage for moving the detection objective along the second longitudinal axis for maintaining the sample in focus.

9. A converted microscope comprising:
a base engaged to a translation stage and a sample stage, the translation stage being adapted to move in the z direction and the sample stage being adapted to move a sample in the x-y-z directions relative to the base;
a mount engaged to the translation stage for moving the mount in the z direction, the mount defining a mount body, the mount further including an excitation objective engaged to one portion of the mount body and oriented along a first longitudinal axis and a detection objective engaged to another portion of the mount body and oriented along a second longitudinal axis, wherein the excitation objective and the detection objective are oriented such that the first longitudinal axis and the second longitudinal axis are oriented in a perpendicular geometric relation to each other, and
a selective plane illumination microscopy (SPIM) arrangement in communication with the excitation objective and the detection objective, the SPIM arrangement including a light source for generating in alternating fashion a first plurality of light sheets through the excitation objective and then a second plurality of light sheets through the detection objective, and vice versa, for producing an excitation in a sample that generates an alternating sequence of a first fluorescent emission detected by the excitation objective and then a second fluorescent emission detected by the detection objective, and vice versa.

10. The converted microscope of claim 9 , further comprising:
a conventional objective arrangement oriented below the sample for detecting the sample when illuminated by the light source.

11. The converted microscope of claim 9 , wherein the mount body defines a first arm engaged to the excitation objective and a second arm engaged to the detection objective.

12. The converted microscope of claim 9 , wherein the SPIM arrangement further includes a lens and mirror arrangement for directing the sheet of light through the excitation objective.

13. A converted microscope comprising:
a base engaged to a translation stage and a sample stage, the translation stage being adapted to move in the z direction and the sample stage being adapted to move a sample in the x-y-z directions relative to the base;
a mount engaged to the translation stage for moving the mount in the z direction, the mount defining a mount body, the mount further including a first objective engaged to one portion of the mount body and oriented along a first longitudinal axis and a second objective engaged to another portion of the mount body and oriented along a second longitudinal axis, wherein the first objective and the second objective are oriented such that the first longitudinal axis and the second longitudinal axis are oriented in a perpendicular geometric relation to each other;
at least one selective plane illumination microscopy (SPIM) arrangement in communication with the first objective and the second objective with the SPIM arrangement including a light source for generating in alternating fashion a first plurality of light sheets through the first objective and then a second plurality of light sheets through the second objective, and vice versa, for producing an excitation in a sample that generates an alternating sequence of a first fluorescent emission detected by the first objective and then a second fluorescent emission detected by the second objective, and vice versa, wherein the first fluorescent emission is oriented in perpendicular orientation relative to the second fluorescent emission.

14. The converted microscope of claim 13 , wherein the first fluorescent emissions are focused onto a first sensor through the second objective and the second fluorescent emissions are focused onto a second sensor through the first objective.

15. The converted microscope of claim 14 , wherein the first sensor and the second sensor communicate with a processor for collecting the alternating sequence of first fluorescent emissions and second fluorescent emissions.

16. The converted microscope of claim 15 , wherein the processor receives a first image from the first sensor and a second image from the second sensor, wherein the first image and the second image are in perpendicular orientation relative to each other.

17. The converted microscope of claim 16 , wherein the processor performs a registration process in which the first image is transposed onto the second image such that the first image is in perpendicular orientation relative to the second image.

18. The converted microscope of claim 17 , wherein the first image defines a first axial resolution and a first lateral resolution and the second image defines a second axial resolution and a second lateral resolution, wherein transposing the first image onto the second image permits the first lateral resolution to be transposed over the second axial resolution as well as the second lateral resolution to be transposed over the first axial resolution such that the first lateral resolution enhances the resolution of the second axial resolution and the second lateral resolution enhances the resolution of the first axial resolution.

19. The converted micr

Assignees

• Shroff Hari
• Wu Yicong
• Rondeau Gary
• Us Health

Inventors

• Shroff Hari
• Wu Yicong
• Rondeau Gary

Classifications

• G02B21/06 Primary

  Means for illuminating specimens · CPC title

• G02B21/0004

  specially adapted for specific applications · CPC title