# US 7,612,350 B2 · Scanning microscope and specimen image obtaining method in which adjacent data obtaining points are not consecutively obtained

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- Number: US 7,612,350 B2
- Document ID: US7612350B2
- Status: Granted
- Kind: B2
- Application number: 11/330,429
- Filed: 2006-01-12
- Published: 2006-07-20
- Granted: 2009-11-03
- Earliest claimed: 2005-01-18
- Assignees: Olympus Corporation
- Inventors: Nobuhiro Takamizawa
- Web page: https://patentgrep.com/patent/US7612350B2

## CPC

- G01N 21/6458: {Fluorescence microscopy (fluorescence microscopes per se G02B21/0076 and G02B21/16)}
- G02B 21/0036: {Scanning details, e.g. scanning stages}
- G02B 21/0052: {Optical details of the image generation}
- G02B 21/0064: {multi-spectral or wavelength-selective arrangements, e.g. wavelength fan-out, chromatic profiling (G02B21/0076 takes precedence)}
- G02B 21/0076: {arrangements using fluorescence or luminescence}
- G02B 21/16: adapted for ultraviolet illumination {; Fluorescence microscopes (G02B21/0076 takes precedence)}
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- G02B: OPTICAL ELEMENTS, SYSTEMS OR APPARATUS

## Abstract

A scanning microscope includes a light source unit that projects laser light, an optical system that converges and applies the laser light onto data obtaining points on a specimen, and a data obtaining order deciding unit that decides a data obtaining order such that adjacent data obtaining points are not consecutive in the data obtaining order. A scanning unit scans the laser light in accordance with the data obtaining order, a detector detects detection light from the data obtaining points, respectively, and a storage unit stores luminance information on the detection light detected by the detector in association with positional information on the data obtaining points, respectively. An image formation unit forms a two-dimensional image based on the stored associated luminance and positional information.

## Claims

### Claim 1

A scanning microscope comprising:
a light source unit that projects laser light;
an optical system that converges and applies the laser light individually onto data obtaining points on a continuous biological specimen;
a data obtaining order deciding unit that decides a data obtaining order such that spatially adjacent data obtaining points are not consecutive in the data obtaining order;
a scanning unit that optically scans the laser light in a two-dimensional plane of the specimen in accordance with the data obtaining order;
a detector that detects detection light from the data obtaining points, respectively;
a storage unit that stores luminance information on the detection light detected by the detector in association with positional information on the data obtaining points, respectively; and
an image formation unit that forms a two-dimensional image based on the stored associated luminance and positional information,
wherein the data obtaining order deciding unit is configured to randomly decide the data obtaining order and check whether the data obtaining order includes sequential two points that are spatially adjacent, and wherein when it is determined that the data obtaining order includes the sequential two points that are spatially adjacent, the data obtaining order deciding unit replaces one of the sequential two points with another point.

### Claim 2 (depends on 1)

The scanning microscope according to claim 1, wherein after the detector detects the detection light from a data obtaining point a plurality of times at predetermined time intervals, the scanning unit moves the laser light to a next data obtaining point, and the image formation unit forms a two-dimensional image for each detection rank by associating each of the luminance information on the detection light at a same rank in the luminance information on the detection light from the data obtaining points detected by the detector with the positional information on the data obtaining points.

### Claim 3 (depends on 1)

The scanning microscope according to claim 1, wherein the detector detects a current value generated in the specimen synchronously with the detection of the detection light from a data obtaining point when the laser light is applied to the data obtaining point.

### Claim 4 (depends on 1)

The scanning microscope according to claim 1, further comprising a laser light control unit that controls at least one of a wavelength and an intensity of the laser light projected from the light source unit, wherein after the detector detects the detection light from a data obtaining point a plurality of times at predetermined time intervals, the scanning unit moves the laser light to a next data obtaining point, and wherein the laser light control unit changes an irradiation condition of the laser light in association with a detection order at each data obtaining point.

### Claim 5 (depends on 4)

The scanning microscope according to claim 4, wherein the irradiation condition of the laser light is at least one of turning on/off of laser light irradiation, intensity of the laser light, and wavelength components of the laser light.

### Claim 6 (depends on 1)

The scanning microscope according to claim 1, wherein the data obtaining order deciding unit is also configured to regularly decide the data obtaining order.

### Claim 7 (depends on 6)

The scanning microscope according to claim 6, wherein the data obtaining order deciding unit is configured to divide a scanning region into plural regions, and decide an order in which the laser light is regularly applied to the plural regions in accordance with a same rule.

### Claim 8

A scanning microscope comprising:
light source means for projecting laser light;
application means for converging and applying the laser light individually onto data obtaining points on a continuous biological specimen;
data obtaining order deciding means for deciding a data obtaining order such that spatially adjacent data obtaining points are not consecutive in the data obtaining order;
scanning means for optically scanning the laser light in a two-dimensional plane of the specimen in accordance with the data obtaining order;
detection means for detecting detection light from the data obtaining points, respectively;
storage means for preserving luminance information on the detection light detected by the detection means in association with positional information on the data obtaining points, respectively; and
image formation means for forming a two-dimensional image based on the stored associated luminance and positional information,
wherein the data obtaining order deciding means is configured to randomly decide the data obtaining order and check whether the data obtaining order includes sequential two points that are spatially adjacent, and wherein when it is determined that the data obtaining order includes the sequential two points that are spatially adjacent, the data obtaining order deciding means replaces one of the sequential two points with another point.

### Claim 9 (depends on 8)

The scanning microscope according to claim 8, wherein after the detection means detects the detection light from a data obtaining point a plurality of times at predetermined time intervals, the scanning means moves the laser light to a next data obtaining point, and the image formation means forms a two-dimensional image for each detection rank by associating each of the luminance information on the detection light at a same rank in the luminance information on the detection light from the data obtaining points detected by the detection means with the positional information on the data obtaining points.

### Claim 10 (depends on 8)

The scanning microscope according to claim 8, wherein the detection means detects a current value generated in the specimen synchronously with the detection of the detection light from a data obtaining point when the laser light is applied to the data obtaining point.

### Claim 11 (depends on 8)

The scanning microscope according to claim 8, further comprising laser light control means for controlling at least one of a wavelength and an intensity of the laser light projected from the light source means, wherein after the detection means detects the detection light from a data obtaining point a plurality of times at predetermined time intervals, the scanning means moves the laser light to a next data obtaining point, and wherein the laser light control means changes an irradiation condition of the laser light in association with a detection order at each data obtaining point.

### Claim 12 (depends on 11)

The scanning microscope according to claim 11, wherein the irradiation condition of the laser light is at least one of turning on/off of laser light irradiation, intensity of the laser light and wavelength components thereof.

### Claim 13 (depends on 8)

The scanning microscope according to claim 8, wherein the data obtaining order deciding means is also configured to regularly decide the data obtaining order.

### Claim 14 (depends on 13)

The scanning microscope according to claim 13, wherein the data obtaining order deciding means is configured to divide a scanning region into plural regions, and decide an order in which the laser light is regularly applied to the plural regions in accordance with a same rule.

### Claim 15

A specimen image obtaining method comprising:
converging and applying laser light individually onto data obtaining points on a continuous biological specimen;
deciding a data obtaining order in which spatially adjacent data obtaining points are not consecutive;
optically scanning the laser light in a two-dimensional plane of the specimen in accordance with the data obtaining order;
detecting detection light from the data obtaining points, respectively;
storing luminance information on the detection light detected from the data obtaining points in association with positional information on the data obtaining points, respectively; and
forming a two-dimensional image based on the stored associated luminance and positional information,
wherein the data obtaining order is decided randomly and it is checked whether the data obtaining order includes sequential two points that are spatially adjacent, and wherein when it is determined that the data obtaining order includes the sequential two points that are spatially adjacent, one of the sequential two points is replaced with another point.

### Claim 16

A scanning microscope comprising:
a light source unit that projects laser light;
an optical system that converges and applies the laser light onto data obtaining points on a continuous biological specimen which contains a compound that causes a photochemical reaction;
a data obtaining order deciding unit that decides a data obtaining order of the data obtaining points such that a second data obtaining point, where data is obtained time-sequentially next to a first data obtaining point, is located away from the first data obtaining point by a distance such that the second data obtaining point is not affected by the photochemical reaction caused by the application of the laser light to the first data obtaining point;
a scanning unit that scans the laser light optically on a two-dimensional plane of the specimen in accordance with the data obtaining order;
a detector that detects detection light from the data obtaining points;
a storage unit that stores luminance information on the detection light detected by the detector in association with positional information on the data obtaining points; and
an image formation unit that forms a two-dimensional image based on the association of the luminance information with the positional information,
wherein the data obtaining order deciding unit is configured to randomly decide the data obtaining order and check whether the data obtaining order includes sequential two points that are spatially adjacent, and wherein when it is determined that the data obtaining order includes the sequential two points that are spatially adjacent, the data obtaining order deciding unit replaces one of the sequential two points with another point.
