# US 9,185,357 B2 · Optical tissue sectioning using full field optical coherence tomography

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- Number: US 9,185,357 B2
- Document ID: US9185357B2
- Status: Granted
- Kind: B2
- Application number: 13/822,852
- Filed: 2011-09-19
- Published: 2013-07-18
- Granted: 2015-11-10
- Earliest claimed: 2010-09-17
- Assignees: Lltech Management
- Applicants: Albert Claude Boccara; Fabrice Harms; Bertrand Le Conte Chrestien De Poly
- Inventors: Albert Claude Boccara; Fabrice Harms; Bertrand Le Conte Chrestien De Poly
- Web page: https://patentgrep.com/patent/US9185357B2

## CPC

- H04N 7/18: Closed-circuit television [CCTV] systems, i.e. systems in which the video signal is not broadcast
- A61B 5/0066: {Optical coherence imaging}
- A61B 5/0068: {Confocal scanning}
- G01B 9/02041: {characterised by particular imaging or detection techniques}
- G01B 9/02091: Tomographic interferometers, e.g. based on optical coherence
- G02B 21/14: affording illumination for phase-contrast observation
- G02B 21/16: adapted for ultraviolet illumination {; Fluorescence microscopes (G02B21/0076 takes precedence)}
- G02B 21/367: {providing an output produced by processing a plurality of individual source images, e.g. image tiling, montage, composite images, depth sectioning, image comparison}
- H04N: PICTORIAL COMMUNICATION, e.g. TELEVISION
- A61B: DIAGNOSIS; SURGERY; IDENTIFICATION (usefulness limited to only animals A61D)
- G01B: MEASURING LENGTH, THICKNESS OR SIMILAR LINEAR DIMENSIONS; MEASURING ANGLES; MEASURING AREAS; MEASURING IRREGULARITIES OF SURFACES OR CONTOURS
- G02B: OPTICAL ELEMENTS, SYSTEMS OR APPARATUS

## Abstract

According to a first aspect, the invention relates to a multimodal optical sectioning microscope (200, 400, 600) for full-field imaging of a volumic and scattering sample comprising: —a full-field OCT system for providing an image of a first section in depth of the sample comprising an illumination sub-system (201, 401, 601) and a full-field imaging interferometer with a detection sub system (208, 408, 608) and an optical conjugation device for optically conjugating the sample and said detection sub system, wherein said optical conjugation device comprises a microscope objective (203, 403, 603), —a supplementary full-field optical sectioning imaging system for providing a fluorescent image of a second section in depth of said sample comprising a structured illumination microscope with an illumination sub system (623), means (421, 422) for generating at the focal plane of said microscope objective of said full-field imaging interferometer a variable spatial pattern illumination and a detection sub system (624), optically conjugated with said focal plane of the microscope objective.

## Claims

### Claim 1

A multimodal optical sectioning microscope for full-field imaging of a volumic and scattering sample comprising:
a full-field Optical Coherence Tomography (OCT) system for providing an image of a first section in depth of the sample comprising a first illumination sub-system and a full-field imaging interferometer, wherein the full-field imaging interferometer comprises:
a first beamsplitter for defining a reference arm and an object arm for receiving the sample, wherein the reference arm comprises a reference mirror and the object arm comprises a microscope objective,
a detection sub system, and
an optical conjugation device for optically conjugating the sample and said detection sub system, wherein said optical conjugation device comprises the microscope objective;
a supplementary full-field optical sectioning imaging system for providing a fluorescent image of a second section in depth of said sample comprising a structured illumination microscope, wherein the structured illumination microscope comprises:
a second illumination sub system separate from the first illumination sub system,
an imaging channel comprises the microscope objective of the full-field imaging interferometer and means for generating at the focal plane of said microscope objective a variable spatial pattern illumination, wherein the imaging channel is optically coupled with the object arm of the full-field imaging interferometer using the first beamplitter, and
a detection sub system, optically conjugated with said focal plane of the microscope objective; and
a second dichroic beamsplitter, wherein light emitted by the first and second illumination sub systems is combined in the object arm using the first beamsplitter and the second dichroic beamsplitter.

### Claim 2 (depends on 1)

The multimodal optical sectioning microscope according to claim 1, wherein the supplementary full-field optical sectioning imaging system and the full-field OCT system have the same detection sub-system.

### Claim 3 (depends on 1)

The multimodal optical sectioning microscope according to claim 1, wherein the supplementary full-field optical sectioning imaging system and the full-field OCT system have two separate detection sub-systems, and wherein the multimodal optical sectioning microscope further comprises a dichroic beamsplitter able to separate light retro diffused by said first section and fluorescent light emitted by said second section.

### Claim 4 (depends on 1)

The multimodal optical sectioning microscope according to claim 1, wherein said means for generating the spatial pattern comprises a mask with variable spatial transmittance or reflectance to be illuminated by the illumination sub-system of said supplementary full-field optical sectioning imaging system and conjugated with the focal plane of said microscope objective.

### Claim 5 (depends on 1)

The multimodal optical sectioning microscope according to claim 1, wherein the second illumination sub-system of said supplementary full-field optical sectioning imaging system is a laser, and said means for generating the spatial pattern comprises a diffuser to be illuminated by said laser and conjugated with the back aperture of said microscope objective.

### Claim 6

A method for acquiring in depth section images of a first section of a volumic and scattering sample using a Full-Field Optical Coherence Tomography (FF-OCT) system comprising a full-field imaging interferometer and in depth section fluorescent images of a second section of said sample using a supplementary full-field optical sectioning imaging system comprising a structured illumination microscope, the method comprising:
acquiring the images of said first section of the sample by:
emitting light using a first illumination sub system,
defining, in the full-field imaging interferometer, using a first beamsplitter, a reference arm and an object arm for receiving the sample, wherein said reference arm comprises a reference mirror and said object arm comprises a microscope objective,
optically conjugating said sample with a detection sub-system of the full-field imaging interferometer using the microscope objective; and
acquiring fluorescence images of said second section of the sample by:
emitting light using a second illumination sub-system, separate from the first illumination sub-system, wherein the light emitted by said first and second illumination sub-systems is combined in the object arm of the full-field imaging interferometer using a second dichroic beamsplitter and said first beamsplitter,
optically coupling using said first beamsplitter an imaging channel of said structured illumination microscope with the object arm of the full-field imaging interferometer, wherein said imaging channel comprises said microscope objective and means for generating at the focal plane of said microscope objective a variable spatial pattern illumination,
generating at the focal plane of said microscope objective of said full-field imaging interferometer a variable spatial pattern illumination,
forming a plurality of images for different spatial pattern illumination on a detection sub-system optically conjugated with the focal plane of said microscope objective, and
processing said plurality of images to obtain said fluorescence images.

### Claim 7 (depends on 6)

A method according to claim 6, further comprising simultaneous acquisition of images from said supplementary full-field optical sectioning imaging system and said FF-OCT system.

### Claim 8 (depends on 7)

A method according to claim 7, wherein simultaneous acquisition is obtained using spectral separation of light retro diffused by said first section and fluorescent light emitted by said second section.

### Claim 9 (depends on 7)

A method according to claim 7, wherein simultaneous acquisition is obtained using a multispectral array detector common for said supplementary full-field optical sectioning imaging system and said FF-OCT system.

### Claim 10 (depends on 7)

A method according to claim 7, wherein simultaneous acquisition is obtained using a single detection sub-system and the discrimination of data coming from said supplementary full-field optical sectioning imaging system, and wherein said FF-OCT system is performed using post processing of data acquired by said detection sub-system.

### Claim 11 (depends on 6)

A method according to claim 6, further comprising displaying said first and second fluorescent images, and displaying merged images from said first and second fluorescent images.

### Claim 12 (depends on 6)

A method according to claim 6, wherein acquiring images of said first section of the sample using the FF-OCT system further comprises acquiring using said imaging interferometer at least a first and a second interferometric images and processing said interferometric images to obtain said in depth section image of said first section.