# US 11,029,486 B2 · Microscope and observation method

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- Number: US 11,029,486 B2
- Document ID: US11029486B2
- Status: Granted
- Kind: B2
- Application number: 16/055,457
- Filed: 2018-08-06
- Published: 2018-11-29
- Granted: 2021-06-08
- Earliest claimed: 2016-02-26
- Assignees: FUJIFILM Corporation
- Applicants: FUJIFILM Corporation
- Inventors: Kenta Matsubara
- Web page: https://patentgrep.com/patent/US11029486B2

## CPC

- G02B 7/28: Systems for automatic generation of focusing signals
- G02B 7/36: using image sharpness techniques {, e.g. image processing techniques for generating autofocus signals (image data processing per se G06T; in cameras having a solid state image sensor H04N23/67)}
- G02B 21/06: Means for illuminating specimens
- G02B 21/241: {Devices for focusing (focusing in general G02B7/28)}
- G02B 21/244: {using image analysis techniques}
- G02B 21/245: {using auxiliary sources, detectors}
- G02B 21/248: {objective (or ocular) turrets}
- G02B 21/26: Stages; Adjusting means therefor
- G02B 21/365: {Control or image processing arrangements for digital or video microscopes (G02B21/361, G02B21/362 take precedence)}
- G03B 13/32: Means for focusing
- H04N 23/67: Focus control based on electronic image sensor signals
- G02B: OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G03B: APPARATUS OR ARRANGEMENTS FOR TAKING PHOTOGRAPHS OR FOR PROJECTING OR VIEWING THEM; APPARATUS OR ARRANGEMENTS EMPLOYING ANALOGOUS TECHNIQUES USING WAVES OTHER THAN OPTICAL WAVES; ACCESSORIES THEREFOR (optical parts of such apparatus G02B; photosensitive materials or processes for photographic purposes G03C; apparatus for processing exposed photographic materials G03D)
- H04N: PICTORIAL COMMUNICATION, e.g. TELEVISION

## Abstract

A microscope includes an illumination light emission unit 10 that emits illumination light, a stage 61 on which a culture container 60 is placed, an objective lens 31 on which the illumination light having passed through the culture container 60 and the stage 61 is incident, a focusing light emission unit 70 that emits focusing light, a reflected light detection unit 75 that detects reflected light due to emission of the focusing light, a distance changing unit 34 that changes a distance between the objective lens 31 and the stage 61, an autofocus control unit 51 that performs autofocus control based on the reflected light, and a focus control information acquisition unit 53 that acquires focus control information including information of the culture container 60, the amount of culture solution C, and the magnification of the objective lens 31.

## Claims

### Claim 1

A microscope, comprising:
an illumination light emission portion that emits illumination light to a container containing a liquid and an observation target;
a container support stage on which the container is placed and which supports the container;
an objective lens on which the illumination light having passed through the container and the container support stage is incident;
a focusing light emission portion that emits focusing light having a wavelength different from that of the illumination light from the container support stage side through the objective lens;
a reflected light detection portion that detects reflected light, which is due to emission of the focusing light from the container support stage side, through the objective lens;
a distance changing mechanism that changes a distance between the objective lens and the container support stage; and
a processor configured to:
perform autofocus control by controlling the distance changing mechanism based on the reflected light detected by the reflected light detection portion; and
acquire focus control information including at least one of information of the container, information of an amount of the liquid, or information of a magnification of the objective lens,
wherein the processor is further configured to change a method of the autofocus control based on the focus control information, and
wherein the processor is further configured to acquire information of a culture period of the observation target as the information of the amount of the liquid.

### Claim 2 (depends on 1)

The microscope according to claim 1,
wherein the processor is further configured to perform switching between first autofocus control, which is for searching for a focal position focused on an observation target placement surface that is a boundary surface between a bottom portion of the container and the observation target, and second autofocus control, which is for searching for a focal position focused on a bottom surface that is a surface of the bottom portion of the container opposite to the observation target placement surface, based on the focus control information.

### Claim 3 (depends on 2)

The microscope according to claim 2,
wherein the processor is further configured to estimate a focal position focused on the observation target placement surface by adding an offset to the focal position searched for by the second autofocus control, and control the distance changing mechanism based on the estimated focal position.

### Claim 4 (depends on 3)

The microscope according to claim 3,
wherein, based on the information of the amount of the liquid, the processor is further configured to perform the first autofocus control in a case where the amount of the liquid is equal to or greater than a liquid threshold value set in advance and perform the second autofocus control in a case where the amount of the liquid is less than the liquid threshold value.

### Claim 5 (depends on 4)

The microscope according to claim 4,
wherein, based on the information of the container, the processor is further configured to perform the first autofocus control in a case where a thickness of the bottom portion of the container is equal to or greater than a thickness threshold value set in advance and perform the second autofocus control in a case where the thickness of the bottom portion of the container is less than the thickness threshold value.

### Claim 6 (depends on 4)

The microscope according to claim 4,
wherein, based on the information of the magnification of the objective lens, the processor is further configured to perform the first autofocus control in a case where the magnification of the objective lens is a relatively high magnification and perform the second autofocus control in a case where the magnification of the objective lens is a relatively low magnification.

### Claim 7 (depends on 3)

The microscope according to claim 3,
wherein, based on the information of the container, the processor is further configured to perform the first autofocus control in a case where a thickness of the bottom portion of the container is equal to or greater than a thickness threshold value set in advance and perform the second autofocus control in a case where the thickness of the bottom portion of the container is less than the thickness threshold value.

### Claim 8 (depends on 3)

The microscope according to claim 3,
wherein, based on the information of the magnification of the objective lens, the processor is further configured to perform the first autofocus control in a case where the magnification of the objective lens is a relatively high magnification and perform the second autofocus control in a case where the magnification of the objective lens is a relatively low magnification.

### Claim 9 (depends on 2)

The microscope according to claim 2,
wherein, based on the information of the amount of the liquid, the processor further configured to perform the first autofocus control in a case where the amount of the liquid is equal to or greater than a liquid threshold value set in advance and perform the second autofocus control in a case where the amount of the liquid is less than the liquid threshold value.

### Claim 10 (depends on 9)

The microscope according to claim 9,
wherein, based on the information of the container, the processor is further configured to perform the first autofocus control in a case where a thickness of the bottom portion of the container is equal to or greater than a thickness threshold value set in advance and perform the second autofocus control in a case where the thickness of the bottom portion of the container is less than the thickness threshold value.

### Claim 11 (depends on 9)

The microscope according to claim 9,
wherein, based on the information of the magnification of the objective lens, the processor is further configured to perform the first autofocus control in a case where the magnification of the objective lens is a relatively high magnification and perform the second autofocus control in a case where the magnification of the objective lens is a relatively low magnification.

### Claim 12 (depends on 2)

The microscope according to claim 2,
wherein, based on the information of the container, the processor is further configured to perform the first autofocus control in a case where a thickness of the bottom portion of the container is equal to or greater than a thickness threshold value set in advance and perform the second autofocus control in a case where the thickness of the bottom portion of the container is less than the thickness threshold value.

### Claim 13 (depends on 12)

The microscope according to claim 12,
wherein, based on the information of the magnification of the objective lens, the processor is further configured to perform the first autofocus control in a case where the magnification of the objective lens is a relatively high magnification and perform the second autofocus control in a case where the magnification of the objective lens is a relatively low magnification.

### Claim 14 (depends on 2)

The microscope according to claim 2,
wherein, based on the information of the magnification of the objective lens, the processor is further configured to perform the first autofocus control in a case where the magnification of the objective lens is a relatively high magnification and perform the second autofocus control in a case where the magnification of the objective lens is a relatively low magnification.

### Claim 15 (depends on 1)

The microscope according to claim 1, further comprising:
a thickness measurement meter that measures a thickness of a bottom portion of the container as the information of the container.

### Claim 16 (depends on 1)

The microscope according to claim 1, further comprising:
an input device that receives a setting input of the focus control information.

### Claim 17 (depends on 1)

The microscope according to claim 1,
wherein the focusing light has a pattern set in advance.

### Claim 18 (depends on 17)

The microscope according to claim 17
wherein the focusing light has a stripe pattern.

### Claim 19

An observation method for observing an image of an observation target by emitting illumination light to a container, which is placed on a container support stage and which contains a liquid and the observation target, and imaging the illumination light, which has passed through the container and the container support stage, through an objective lens, the method comprising:
emitting focusing light having a wavelength different from that of the illumination light from the container support stage side through the objective lens;
in detecting reflected light due to emission of the focusing light and performing autofocus control by changing a distance between the objective lens and the container support stage based on the detected reflected light, acquiring focus control information including at least one of information of the container, information of an amount of the liquid, or information of a magnification of the objective lens;
acquiring information of a culture period of the observation target as the information of the amount of the liquid, and
changing a method of the autofocus control based on the acquired focus control information.
