System and method of edge-illumination microscopy

Patent metadata

Field	Value
Publication number	US-10031326-B2
Application number	US-201414914844-A
Country	US
Kind code	B2
Filing date	Aug 27, 2014
Priority date	Aug 28, 2013
Publication date	Jul 24, 2018
Grant date	Jul 24, 2018

Abstract

According to one aspect, the invention concerns a method for microscopy of a thick sample arranged on a sample support, with edge-illumination of the sample. The method comprises, in particular, emitting at least one illumination beam (1), forming, from the illumination beam, an illumination surface, focusing the illumination surface in the sample by means of a microscope lens (120) and deflecting the illumination surface originating from the microscope lens, in order to form a transverse illumination surface, located in a plane substantially perpendicular to the optical axis of the microscope lens. The method further comprises forming, by means of said microscope lens (120), the image of an area of the sample illuminated by the transverse illumination surface on a detection surface (131) of a detection device (130), scanning the illumination beam, allowing the transverse illumination surface to move along the optical axis of the microscope lens, and superimposing the object imaging surface and the transverse illumination surface, by focusing means comprising means separate from the means for the relative axial movement of the microscope lens and the sample.

First claim

The invention claimed is:

1. A microscopy system for a thick sample with edge-illumination of the sample, comprising:
a sample holder;
a detection pathway comprising:
a microscope lens of given optical axis and exit pupil;
a detection device comprising a detection surface optically conjugate with an object imaging surface in the object space of the microscope lens;
means for relative axial displacement of the microscope lens and of the sample holder;

a sample illumination pathway comprising:
at least one emission source of an illumination beam;
means for forming, from the illumination beam, an illumination surface;
said microscope lens;
deflection means making it possible to deflect the illumination surface in the object space at the output of the microscope lens, to form a transverse illumination surface, situated in a plane substantially at right angles to the optical axis of the microscope lens;
scanning means for an angular scanning of the illumination beam, allowing a displacement of the transverse illumination surface along the optical axis of the microscope lens;

the microscopy system further comprising:
optical means for varying the focusing of the illumination beam, making it possible to laterally center the transverse illumination surface on the object imaging surface, wherein said optical means are separate from the relative axial displacement means; and
relay optics configured for optically conjugating a plane of a pupil of said optical means for varying the focusing of the illumination beam with a plane of the exit pupil of the microscope lens.

2. The microscopy system as claimed in claim 1 , in which the optical means for varying the focusing of the illumination beam comprise an optical system with fixed power coupled to an optical system with variable focal length.

3. The microscopy system as claimed in claim 1 , further comprising a wavefront spatial modulation device, situated in the detection pathway, in which the wavefront spatial modulation device allows for the correction of at least a part of the optical defects between the sample and the detection surface.

4. The microscopy system as claimed in claim 1 , in which the deflection means are secured to the sample holder.

5. The microscopy system as claimed in claim 1 , in which the means for forming the illumination surface comprise means for generating a light pencil and means for scanning the light pencil.

6. The microscopy system as claimed in claim 1 , in which the means for forming the illumination surface comprise a cylindrical lens.

7. The microscopy system as claimed in claim 1 , in which said pupil of the optical means for varying the focusing of the illumination beam is located in a plane optically conjugated with a plane comprising an axis of rotation of said scanning means.

8. An edge-illumination device adapted to a microscope comprising a microscope lens with an object imaging surface, means for relative axial displacement of a sample holder with the microscope lens and a detector with a detection surface optically conjugate with the object imaging surface, the illumination device comprising:
at least one emission source of an illumination beam;
means for forming, from the illumination beam, an illumination surface;
a sample holder and deflection means secured to the sample holder, the deflection means being intended to deflect the illumination surface at the output of the microscope lens, to form a transverse illumination surface, situated in a plane substantially at right angles to the optical axis of the microscope lens;
illumination beam scanning means allowing a displacement of the transverse illumination surface along the optical axis of the microscope lens;
optical means for varying the focusing of the illumination beam, making it possible to laterally center the transverse illumination surface on the object imaging surface, wherein said optical means are separate from the relative axial displacement means; and
relay optics configured for optically conjugating a plane of a pupil of said optical means for varying the focusing of the illumination beam with a plane of the exit pupil of the microscope lens.

9. A microscopy method for a thick sample arranged on a sample holder, with edge-illumination of the sample, comprising:
the emission of at least one illumination beam;
the formation, from the illumination beam, of an illumination surface;
the focusing of the illumination surface in the sample by means of a microscope lens of given optical axis and exit pupil and the deflection of the illumination surface deriving from the microscope lens allowing the formation of a transverse illumination surface, situated in a plane substantially at right angles to the optical axis of the microscope lens;
the formation, by means of said microscope lens, of the image of a zone of the sample illuminated by the transverse illumination surface on a detection surface of a detection device;
the scanning of the illumination beam allowing a displacement of the transverse illumination surface along the optical axis of the microscope lens;
the variation of the focusing of the illumination beam, allowing for a lateral centering of the transverse illumination surface on the object imaging surface, conjugate in the object space of the microscope lens with the detection surface, by optical means separate from the relative axial displacement means; and
the optical conjugation, by relay optics, of a plane of a pupil of said optical means for varying the focusing of the illumination beam with a plane of the exit pupil of the microscope lens.

10. The microscopy method as claimed in claim 9 , further comprising the correction of at least a part of the optical defects between the sample and the detection surface using a wavefront spatial modulation device.

11. The microscopy method as claimed in claim 9 , in which the formation, from the illumination beam, of an illumination surface comprises the generation of a light pencil and the scanning of the light pencil.

Assignees

• Imagine Optic
• Centre National De La Rech Scientifique—Cnrs
• Univ De Bordeaux Segalen
• Centre Nat Rech Scient
• Univ Bordeaux

Inventors

• Levecq Xavier
• Viasnoff Virgile
• Sibarita Jean-Baptiste
• Studer Vincent
• Galland Rémi

Classifications

• G01N2201/10

  Scanning · CPC title

• G01N21/6428

  Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" (in vivo A61B5/00; immunoassay G01N33/53) · CPC title

• G02B21/06 Primary

  Means for illuminating specimens · CPC title

• G01N2201/06113

  Coherent sources; lasers · CPC title

• G02B27/0068

  having means for controlling the degree of correction, e.g. using phase modulators, movable elements (controlling the phase of light using moving or deformable elements G02B26/06) · CPC title