3D localisation microscopy and 4D localisation microscopy and tracking methods and systems

Patent metadata

Field	Value
Publication number	US-9523846-B2
Application number	US-201113825794-A
Country	US
Kind code	B2
Filing date	Sep 26, 2011
Priority date	Sep 24, 2010
Publication date	Dec 20, 2016
Grant date	Dec 20, 2016

Abstract

A 3D localization microscopy system, 4D localization microscopy system, or an emitter tracking system arranged to cause a phase difference between light passing to or from one part of the objective relative to light passing to or from another part of the objective, to produce a point emitter image which comprises two lobes, a separation between which is related to the position of the emitter relative to the objective of the imaging system, and in the 4D system a further property of which image or of said light to or from the objective is related to another location independent property of the emitter.

First claim

The invention claimed is:

1. A 4D localization microscopy or emitter tracking system arranged to cause a phase difference between light passing to or from one part of an objective relative to light passing to or from another part of the objective, to produce a point emitter image which comprises two lobes, a separation between the two lobes being related to the position of the emitter relative to the objective of the imaging system, and a further property of which image or of said light to or from the objective is related to another location independent property of the emitter wherein the further property is a separation between the image lobes.

2. A system according to claim 1 , wherein the system is arranged to cause said phase difference between light passing to or from one part of the objective relative to light passing to or from another part of the objective to produce a point emitter image which comprises two image lobes, wherein in a position of an emitter relative to the objective causing negative defocus, the two image lobes are separated from each other in first opposite directions substantially along an axis, and in a position of an emitter relative to the objective causing positive defocus, the two image lobes are separated from each other substantially along said axis in second opposite directions.

3. A system according to claim 1 , wherein the further property is a separation between the image lobes in an axis at an angle to the axis of the separation between the image lobes related to the position of the emitter.

4. A system according to claim 1 , wherein the further property of the image is a separation between the image lobes in an axis substantially orthogonal to the axis of the separation between the image lobes related to the position of the emitter.

5. A system according to claim 1 , wherein the location independent property of the emitter is any one or more of a spectral content, lifetime, or polarization of light from the emitter.

6. A system according to claim 1 , arranged so that in an in focus position of an emitter relative to the objective the two image lobes are substantially not separated from each other along the axis of separation between the image lobes related to the position of the emitter.

7. A system according to claim 6 , arranged so that in said in focus position of an emitter relative to the objective the two image lobes are separated from each other substantially orthogonal to the axis of separation of the image lobes related to the position of the emitter.

8. A 4D localization microscopy or emitter tracking system arranged to cause a phase difference between light passing to or from one part of an objective relative to light passing to or from another part of the objective, to produce a point emitter image which comprises two lobes, a separation between the two lobes being related to the position of the emitter relative to the objective of the imaging system, and a further property of which image or of said light to or from the objective is related to another location independent property of the emitter, and further comprising a refractive phase change device arranged to cause said phase difference between light passing to or from one part of the objective relative to light passing to or from another part of the objective wherein the refractive phase change device comprises a phase ramp.

9. A system according to claim 8 wherein the phase ramp is a linear phase ramp.

10. A system according to claim 8 , wherein the phase change device comprises a single phase ramp arranged to cause said phase difference to light passing to or from one part of the objective relative to light passing to or from another part of the objective.

11. A system according to claim 8 wherein the phase change device comprises at least two phase ramps, at least one phase ramp arranged to cause a phase change to light passing to or from one part of the objective and at least another phase ramp arranged to cause a phase change to light passing to or from another part of the objective.

12. A system according to claim 11 wherein the two phase ramps have different phase change gradients.

13. A system according to claim 11 , wherein the two phase ramps have opposite phase change gradients.

14. A system according to claim 11 wherein the two phase ramps have equal but opposite phase change gradients.

15. A system according to claim 8 wherein the phase change device comprises multiple linear ramps in a sawtooth configuration.

16. A 4D localization microscopy or emitter tracking system arranged to cause a phase difference between light passing to or from one part of an objective relative to light passing to or from another part of the objective, to produce a point emitter image which comprises two lobes, a separation between the two lobes being related to a position of an emitter relative to the objective of the imaging system, and a further property of which image or of said light to or from the objective is related to another location independent property of the emitter comprising a reflective phase change device arranged to cause said phase difference between light passing to or from one part of the objective relative to light passing to or from another part of the objective wherein the reflective phase change device comprises at least two reflective phase ramps, at least one arranged to cause a phase change to light passing to or from one part of the objective and at least another arranged to cause a phase change to light passing to or from another part of the objective.

17. A system according to claim 16 wherein the reflective phase change devices comprise reflective surfaces at a fixed relative angle.

18. A system according to claim 16 wherein the reflective phase change devices comprise reflective surfaces at an adjustable relative angle.

19. A system according to claim 16 , wherein the reflective phase change devices are arranged to cause a linearly varying phase difference.

20. A system according to claim 16 wherein the reflective phase change device comprises multiple linear reflective phase ramps in a sawtooth configuration.

21. A 4D localization microscopy or emitter tracking system arranged to cause a phase difference between light passing to or from one part of an objective relative to light passing to or from another part of the objective, to produce a point emitter image which comprises two lobes, a separation between the two lobes being related to a position of an emitter relative to the objective of the imaging system, and a further property of which image or of said light to or from the objective is related to another location independent property of the emitter;
comprising a reflective phase change device arranged to cause said phase difference between light passing to or from one part of the objective relative to light passing to or from another part of the objective;
wherein the reflective phase change device comprises a deformable mirror.

22. A 4D localization microscopy or emitter tracking system arranged to cause a phase difference between light passing to or from one part of an objective relative to light passing to or from another part of the objective, to produce a point emitter image which comprises two lobes, a separation between the two lobes being related to a position of an emitter relative to the objective of the imaging system, and a further property of which image or of said light to or from the objective is related to another location independent property

Assignees

• Soeller Christian
• Baddeley David Michael
• Cannell Mark Bryden
• Zeiss Carl Microscopy Gmbh

Inventors

• Soeller Christian
• Baddeley David Michael
• Cannell Mark Bryden

Classifications

• G02B21/365 Primary

  Control or image processing arrangements for digital or video microscopes (G02B21/361, G02B21/362 take precedence) · CPC title

• G01N21/6458 Primary

  Fluorescence microscopy (fluorescence microscopes per se G02B21/0076 and G02B21/16) · CPC title

• G02B21/0004

  specially adapted for specific applications · CPC title

• H04N17/002

  for television cameras · CPC title

• H04N13/204

  using stereoscopic image cameras (stereoscopic photography G03B35/00) · CPC title