Sample-image acquisition apparatus, sample-image acquisition method and sample-image acquisition program

• US 8,922,637 B2
• Filed: 02/23/2011
• Issued: 12/30/2014
• Est. Priority Date: 03/02/2010
• Status: Expired due to Fees

• Current Assignee: Sony Corporation (Sony Group Corp.)
• Original Assignee: Sony Corporation (Sony Group Corp.)
• Inventors: Kishima, Koichiro
• Primary Examiner(s): Patel, Jay
• Assistant Examiner(s): MIKESKA, NEIL R
• Application Number:
  US13/033,019
• Publication Number:
  US 20110216182A1
• Time in Patent Office: 1,406 Days
• Field of Search: 348/79, 382/128, 250/201.3
• US Class Current: 348/79
• CPC Class Codes: G02B 21/0088 Inverse microscopes
• CPC Class Codes: G02B 21/26 Stages; Adjusting means therefor
• CPC Class Codes: G02B 21/365 Control or image processing arrangements for digital or video microscopes G02B21/361, G02B21/362 take precedence
• CPC Class Codes: 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
• CPC Class Codes: G06T 7/571 from focus
• CPC Class Codes: G06V 20/69 Microscopic objects, e.g. biological cells or cellular parts
• CPC Class Codes: H04N 23/71 Circuitry for evaluating the brightness variation
• CPC Class Codes: H04N 7/18 Closed-circuit television [CCTV] systems, i.e. systems in which the video signal is not broadcast

Abstract

A sample-image acquisition apparatus includes: an image taking device; a movement control section; and a sample-image acquisition section.

Claims

1. A sample-image acquisition apparatus comprising:

an image taking device on which an image of a sample is created after being enlarged by an objective lens;

a movement control section configured to move a relative position, which is movable in a plane direction perpendicular to an optical-axis direction as a position relative between said image of said sample and said image taking device used for creating said image of said sample after being enlarged by said objective lens having a focal point movable within a range including said sample, in said plane direction changed for each position of said focal point in said optical-axis direction while moving said focal point in said optical-axis direction within said range; and

a sample-image acquisition section configured to acquire a sample image by exposing said image taking device to said sample while said movement control section is moving said relative position in a circular arc at a constant speed in said plane direction and moving said focal point in said optical-axis direction such that a fluorescent marker of the sample is in focus for at least a portion of the exposure and subsequently blurred for another portion of the exposure.

2. The sample-image acquisition apparatus according to claim 1, wherein, while moving said focal point of said objective lens in said optical-axis direction within said range including said sample, said movement control section moves a relative position, which is movable in said plane direction as a position relative between a sample-member image created on said image taking device as an image of a member of said sample after being enlarged by said objective lens and said image taking device, along circumferences of circles having the same center point from start points different from each other.

3. The sample-image acquisition apparatus according to claim 2, wherein:

said movement control section moves said focal point of said objective lens in said optical-axis direction within said range including said sample at said center point of said circle having a circumference, along which said movement control section moves said relative position in said plane direction; and

while said movement control section is moving said focal point of said objective lens in said optical-axis direction, said sample-image acquisition section acquires an image of said sample by exposing said image taking device to said sample.

4. The sample-image acquisition apparatus according to claim 1, wherein, while moving said focal point of said objective lens in said optical-axis direction within said range including said sample, said movement control section moves a relative position, which is movable in said plane direction as a position relative between a sample-member image created on said image taking device as an image of a member of said sample after being enlarged by said objective lens and said image taking device, along straight lines in order to change said plane direction.

5. The sample-image acquisition apparatus according to claim 1, said sample-image acquisition apparatus further having a computation section configured to compute a position of said sample in said optical-axis direction on the basis of a movement locus traced by a sample-member image of said sample image on said image taking device.

8. The sample-image acquisition apparatus according to claim 1, wherein the sample image indicates a depth of the fluorescent marker in said sample.

9. The sample-image acquisition apparatus according to claim 1, wherein while said image taking device is exposed to said sample, said movement control section moves the focal point in the optical-axis direction from a top of the sample to a bottom of the sample.

10. The sample-image acquisition apparatus according to claim 1, wherein the movement control section moves at least one of the focal point and the relative position such that the fluorescent marker associated with the sample is (i) blurred during a first portion of the exposure, (ii) in focus during a second portion of the exposure, and (iii) blurred during a third portion of the exposure, the first portion occurring before the second portion and the third portion and the second portion occurring before the third portion.

11. The sample-image acquisition apparatus according to claim 1, wherein the movement control section moves at least one of the focal point and the relative position to distinguish a three-dimensional location of the fluorescent marker from a three-dimensional location of an adjacent fluorescent marker.

12. The sample-image acquisition apparatus according to claim 1,

wherein said movement control section is configured to move said relative position in the circular arc in said plane direction according to equation 1 and equation 2,

wherein equation 1 is equal to X=X0+L0(sin(ω

T)) and equation 2 is to Y=Y0+L0(cos(ω

T)), and

wherein X is a lateral position in said plane direction, Y is a longitudinal position in said plane direction, X0 and Y0 correspond to a reference position within said plane direction, L0 is a radius of the circular arc, and T is a movement time.

13. The sample-image acquisition apparatus according to claim 1,

wherein said movement control section is configured to move said relative position in the circular arc in said plane direction according to equation 1 and equation 2,

wherein equation 1 is equal to X=X0+L0(sin(ω

T+90°

)) and equation 2 is to Y=Y0+L0(cos(ω

T+90°

)), and

wherein X is a lateral position in said plane direction, Y is a longitudinal position in said plane direction, X0 and Y0 correspond to a reference position within said plane direction, L0 is a radius of the circular arc, and T is a movement time.

6. A sample-image acquisition method comprising:

a movement control step of moving a relative position, which is movable in a plane direction perpendicular to an optical-axis direction as a position relative between an image of a sample and an image taking device used for creating said image of said sample after being enlarged by an objective lens having a focal point movable within a range including said sample, in said plane direction changed for each position of said focal point in said optical-axis direction while moving said focal point in said optical-axis direction within said range; and

a sample-image acquisition step of acquiring a sample image by exposing said image taking device to said sample while said movement control step is being carried out to move said relative position in a circular arc at a constant speed in said plane direction and move said focal point in said optical-axis direction such that a fluorescent marker of the sample is in focus for at least a portion of the exposure and subsequently blurred for another portion of the exposure.

7. A non-transitory computer readable storage medium storing a sample-image acquisition program to be executed by a computer to perform:

a movement control step of moving a relative position, which is movable in a plane direction perpendicular to an optical-axis direction as a position relative between an image of a sample and an image taking device used for creating said image of said sample after being enlarged by an objective lens having a focal point movable within a range including said sample, in said plane direction changed for each position of said focal point in said optical-axis direction while moving said focal point in said optical-axis direction within said range; and

a sample-image acquisition step of acquiring a sample image by exposing said image taking device to said sample while said movement control step is being carried out to move said relative position in a circular arc at a constant speed in said plane direction and move said focal point in said optical-axis direction such that a fluorescent marker of the sample is in focus for at least a portion of the exposure and subsequently blurred for another portion of the exposure.