WO2019003410A1 - Cell image acquisition device and cell image acquisition method - Google Patents

Cell image acquisition device and cell image acquisition method Download PDF

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Publication number
WO2019003410A1
WO2019003410A1 PCT/JP2017/024112 JP2017024112W WO2019003410A1 WO 2019003410 A1 WO2019003410 A1 WO 2019003410A1 JP 2017024112 W JP2017024112 W JP 2017024112W WO 2019003410 A1 WO2019003410 A1 WO 2019003410A1
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imaging
imaging unit
interest
unit
image
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PCT/JP2017/024112
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French (fr)
Japanese (ja)
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朗 松下
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オリンパス株式会社
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Priority to PCT/JP2017/024112 priority Critical patent/WO2019003410A1/en
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/34—Measuring or testing with condition measuring or sensing means, e.g. colony counters
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • H—ELECTRICITY
    • H04—ELECTRIC COMMUNICATION TECHNIQUE
    • H04N—PICTORIAL COMMUNICATION, e.g. TELEVISION
    • H04N23/00—Cameras or camera modules comprising electronic image sensors; Control thereof
    • H04N23/60—Control of cameras or camera modules
    • H—ELECTRICITY
    • H04—ELECTRIC COMMUNICATION TECHNIQUE
    • H04N—PICTORIAL COMMUNICATION, e.g. TELEVISION
    • H04N23/00—Cameras or camera modules comprising electronic image sensors; Control thereof
    • H04N23/70—Circuitry for compensating brightness variation in the scene

Definitions

  • the present invention relates to a cell image acquisition apparatus and a cell image acquisition method.
  • An observation target is identified in image data corresponding to a wide area of a biological sample acquired by low magnification imaging, a high magnification image is acquired by high magnification imaging for the identified observation target, and the acquired high magnification image is tied
  • An observation device is known which rings and constructs a wide range of high magnification images of a biological sample (see, for example, Patent Document 1).
  • Patent No. 5516108 gazette
  • the present invention has been made in view of the above-described circumstances, and it is possible to obtain a high-contrast image of a wide area of a culture vessel in a short time and to appropriately manage the quality of a biological sample It aims at providing an acquisition device and a cell image acquisition method.
  • One aspect of the present invention is a first imaging unit for imaging light from cells in a culture vessel, a second imaging unit for imaging with high resolution and a narrower visual field range than the first imaging unit, and 1.
  • a region-of-interest setting unit for setting one or more regions of interest narrower than the field of view range of the first imaging unit determined by analyzing the first image acquired by the imaging unit and determining that there is a cell;
  • An imaging condition setting unit configured to set an imaging condition by the first imaging unit based on a second image acquired by the second imaging unit for each of the regions of interest set by the unit;
  • the first imaging unit is configured to acquire a third image based on the imaging condition set by the user.
  • the first image of the culture container is acquired by photographing the light from the cells in the culture container by the first imaging unit
  • the first image is analyzed by the region of interest setting unit, and the cells are One or more regions of interest determined to exist are set. Then, a second image is acquired for each region of interest by the second imaging unit.
  • the imaging condition setting unit can appropriately set the imaging condition by the first imaging unit, and the third imaging unit acquires the third image by the first imaging unit based on the set imaging condition.
  • the high-contrast image of the region can be acquired in a short time to properly manage the cell quality.
  • the photographing condition setting unit may set the in-focus position by the first imaging unit.
  • the focusing range is narrow and the contrast value is strictly evaluated by acquiring by the second imaging unit that performs imaging with high resolution and narrower than the visual field range by the first imaging unit.
  • An appropriate in-focus position by the first imaging unit can be set using an image that can be captured. Thereby, high-contrast images of a wide area of the culture vessel can be obtained in a short time, and the cell quality can be properly managed.
  • the photographing condition setting unit may set an exposure condition by the first imaging unit.
  • the focusing range is narrow and the contrast value is strictly evaluated by acquiring by the second imaging unit that performs imaging with high resolution and narrower than the visual field range by the first imaging unit.
  • An appropriate exposure condition by the first imaging unit can be set using an image that can be set. Thereby, high-contrast images of a wide area of the culture vessel can be obtained in a short time, and the cell quality can be properly managed.
  • the region of interest setting unit divides the first image acquired by the first imaging unit into a plurality of divided regions, and the number or number set in advance is set in each of the divided regions.
  • the number of regions of interest may be set equal to or less than the maximum number.
  • the first optical lens and the second optical lens, the exchange mechanism for exchanging the first optical lens and the second optical lens, and the collection by the first optical lens or the second optical lens And an imaging device configured to capture the emitted light, wherein the first imaging unit is configured by a combination of the first optical lens and the imaging device, and the second imaging unit is the second optical lens and the second imaging unit.
  • You may be comprised by the combination with an image pick-up element.
  • the zoom optical system includes the zoom optical system and an image pickup element for photographing light condensed by the zoom optical system, and the first image pickup unit arranges the zoom optical system at a first zoom position.
  • the second imaging unit may be configured by disposing the zoom optical system at a second zoom position.
  • the other aspect of this invention is a 1st imaging
  • photography step acquired A region of interest setting step of analyzing one image to set one or more regions of interest narrower than the first visual field range determined to have the cells present, and the respective regions of interest set in the region of interest setting step A shooting condition based on a second shooting step of shooting at a resolution higher than that of the first shooting step to obtain one or more second images, and the one or more second images obtained in the second shooting step
  • ADVANTAGE OF THE INVENTION According to this invention, it is effective in the ability to acquire the high contrast image of the wide area
  • FIG. 1 It is a whole block diagram which shows the cell image acquisition apparatus which concerns on one Embodiment of this invention. It is a longitudinal cross-sectional view which shows the base part of the cell image acquisition apparatus of FIG. It is a top view which shows the base part of the cell image acquisition apparatus of FIG. It is a flowchart explaining the setting method of the exposure conditions in the cell image acquisition apparatus of FIG. It is a flowchart explaining the cell image acquisition method using the cell image acquisition apparatus of FIG. It is a figure which shows an example which sets a part of interest area
  • FIG. 1 It is a figure which shows the other example which sets a part of interest area
  • the cell image acquisition apparatus 1 is an apparatus for acquiring an image of the cell X accommodated and cultured in the culture container 2, and as shown in FIGS. 1 to 3, the visual field range and
  • the first imaging unit 3 and the second imaging unit 4 having different resolutions, and the moving mechanism 5 for horizontally moving the imaging units 3 and 4 are provided.
  • the base unit 10 is configured by the first imaging unit 3, the second imaging unit 4, and the moving mechanism 5.
  • the first imaging unit 3 is, for example, a line sensor including a plurality of pixels which are moved in one direction by the moving mechanism 5 and arranged in a direction crossing the moving direction by the moving mechanism 5. By sequentially acquiring line-shaped images while being driven, a first image of a wide first field of view over substantially the entire bottom surface of the culture vessel 2 is acquired. Note that the first visual field range may not necessarily cover substantially the entire bottom surface of the culture vessel 2. Further, the first imaging unit 3 is not limited to the line sensor, and any other imaging unit may be adopted.
  • the second imaging unit 4 includes, for example, a solid-state imaging device (not shown) that acquires a second image of high resolution from the first imaging unit 3 in a second visual field range narrower than the first imaging unit 3.
  • a solid-state imaging device (not shown) that acquires a second image of high resolution from the first imaging unit 3 in a second visual field range narrower than the first imaging unit 3.
  • the first imaging unit 3 and the second imaging unit 4 are provided with a focusing mechanism and an exposure adjustment mechanism not shown.
  • the focusing mechanism is, for example, a mechanism that moves one or more lenses in the optical axis direction.
  • the exposure adjustment mechanism is, for example, a shutter.
  • the moving mechanism 5 is rotated by an X-axis drive motor 12 that rotates the X-axis ball screw 11, the X-axis ball screw 11, and the X-axis drive motor 12 as shown in FIGS. 2 and 3.
  • a Y-axis drive screw 13 which moves on the X-axis ball screw 11 in one direction (X-direction)
  • a Y-axis ball screw 14 which is disposed orthogonal to the X-axis ball screw 11 and is rotated by the Y-axis drive motor 13
  • a slider 15 moving in one direction (Y direction) on the Y-axis ball screw 14 rotated by the Y-axis drive motor 13.
  • the first imaging unit 3 and the second imaging unit 4 are fixed to the slider 15, and the slider 15 horizontally moves in two directions (XY directions) orthogonal to each other.
  • the cell image acquisition apparatus 1 has the region of interest setting unit 6 connected to the first imaging unit 3 and the imaging condition setting connected to the second imaging unit 4.
  • a control unit 8 for controlling the unit 7, the first imaging unit 3, the second imaging unit 4 and the moving mechanism 5, and a display unit 9 for displaying the third image finally acquired by the first imaging unit 3; Have.
  • a storage unit for storing the third image may be provided instead of or in addition to the display unit 9.
  • the region of interest setting unit 6 analyzes the first image over the wide first field range acquired by the first imaging unit 3 to determine whether or not the cell X is present, and it is determined that the cell X is present.
  • the region of interest is set as a region of interest (ROI) sufficiently smaller than the first visual field range. Specifically, the center coordinates of the set region of interest are stored. If there are a plurality of regions determined that the cell X is present, a plurality of regions of interest are set, and if there are regions determined that the cell X is present over a wider range than the regions of interest, a plurality of regions are determined. Regions of interest will be set adjacent to each other.
  • ROI region of interest
  • the imaging condition setting unit 7 obtains the optimal focusing position and the optimal exposure condition (exposure condition) in each region of interest, using the second image for each region of interest acquired by the second imaging unit. There is. Specifically, with regard to the in-focus position, imaging is performed while adjusting the in-focus position by the second imaging unit 4 in each region of interest, and the in-focus range is detected. When the detected in-focus range does not exceed the depth of field of the first imaging unit 3, the center position of the in-focus range is set as the in-focus position, and the in-focus range is the first imaging unit 3. When the depth of field is exceeded, the focusing range is divided by the depth of field and the focusing position is set at the center position of each depth of field.
  • step S20 it is judged whether or not the exposure time is adjusted (step S20), and whether or not the image is allowed to be saturated regardless of the adjustment of the exposure time Is determined (steps S21 and S22).
  • the exposure time is adjusted, and when saturation of the image is allowed, it is set to shoot at the average exposure time (step S23), and the process is ended.
  • the exposure time is adjusted, and when the saturation of the image is not permitted, it is set to shoot at least twice or more with the exposure time from the minimum to the maximum (step S24), and the process is ended. If the exposure time is not adjusted and saturation of the image is allowed, shooting is performed with the illumination light amount corresponding to the average exposure time (step S25), and the process is ended.
  • step S26 shooting is performed with the illumination light amount corresponding to the minimum to maximum exposure time (step S26), and the process is ended.
  • an appropriate shutter speed is detected in each region of interest using the automatic exposure function of the second imaging unit 4.
  • the exposure condition can be adjusted by the shutter speed, the light amount of the illumination light emitted from the illumination light source (not shown), the opening degree of the aperture, or the like.
  • the shutter speed is set to the average value of the detected shutter speeds in the pixel array direction of the first imaging unit 3 and the first imaging unit 3 performs imaging while changing the shutter speed in the moving direction by the moving mechanism 5 Is set as
  • a cell image acquisition method using the cell image acquisition device 1 according to the present embodiment configured as described above will be described below.
  • the control unit 8 controls the focus of the first imaging unit 3.
  • the first image of the first field of view over the entire bottom surface of the culture container 2 is obtained while setting the position slightly above the bottom surface of the culture container 2 and moving the first imaging unit 3 by the moving mechanism 5 (first Shooting) (first shooting step S1).
  • the acquired first image is sent to the region of interest setting unit 6, and analysis of whether or not the cell X is present is performed, and the cell X is present. Then, the recognized region is set as a region of interest (region of interest setting step S2).
  • the coordinates (for example, center coordinates) of each set region of interest are stored (step S3).
  • the control unit 8 After the coordinates of all the regions of interest are stored, the control unit 8 causes the moving mechanism 5 to move the second imaging unit 4 so that the second view range of the second imaging unit 4 matches any of the regions of interest. (Step S4). In this state, the control unit 8 causes the second imaging unit 4 to acquire a second image (second imaging) while changing the in-focus position of the second imaging unit 4 in the optical axis direction (second imaging step S5).
  • the imaging condition setting unit 7 determines a range in which the cell X exists in the optical axis direction as a focusing range, and a shutter that provides appropriate exposure in the focusing range The speed is detected as an exposure condition and stored (shooting condition setting step S6).
  • step S7 It is determined whether the in-focus range and the exposure condition have been detected for all the regions of interest (step S7), and if not completed, the process from step S4 is repeated.
  • the imaging condition setting unit 7 obtains the in-focus range of the entire culture vessel 2 from the in-focus regions of all the areas of interest (step S8).
  • step S9 it is determined whether the in-focus range of the entire culture container 2 exceeds the depth of field by the first imaging unit 3 (step S9).
  • the focusing range is divided into a plurality of blocks according to the depth of field by the first imaging unit 3, and the focusing position and the exposure condition are set for each block (step S10). If not exceeded, the process of step S11 described later is executed.
  • step S11 the average value of the shutter speeds calculated for the plurality of regions of interest is different for each imaging position different in the movement direction by the moving mechanism 5. The shutter speed is set.
  • the control unit 8 causes the first imaging unit 3 to operate again using the imaging conditions set by the imaging condition setting unit 7, and the first imaging unit 3 acquires the third image of the first visual field range ( The third shooting) is performed (third shooting step S11).
  • the third imaging step S11 when imaging conditions of a plurality of blocks are set in step S10, acquisition of the third image is performed a number of times equal to the number of blocks according to the in-focus position and exposure condition for each block. It will be.
  • the region of interest is based on the wide-area first image acquired by the first imaging unit 3 having the wide-range first visual field range.
  • imaging conditions are set based on the second image acquired by the second imaging unit 4 having a narrower field of view than the first imaging unit 3 and high resolution for each region of interest.
  • the imaging condition by the first imaging unit 3 can be appropriately set by the second image in which the focusing range is narrow and the evaluation of the contrast value can be strictly performed.
  • the in-focus range detected by the second imaging unit 4 exceeds the depth of field of the first imaging unit 3
  • the in-focus range is divided and divided into a plurality of times to divide the third image. Since the cells X are obtained, even if the cell X grows and the size in the height direction is increased, there is an advantage that the range in which the cell X is present can be observed without being missed. For example, even in the case where the bottom surface of the culture container 2 is not flat but has a height difference, etc., it is possible to observe the cell X without losing it.
  • the second image is acquired for all the detected regions of interest, but instead of this, the imaging conditions may be set using only a part of the regions of interest Good.
  • the imaging conditions may be set using only a part of the regions of interest Good.
  • the first image is divided into a plurality of regions R1, R2, R3, R4, and each region R1, R2, R3 is , R4 may be used for setting the imaging conditions.
  • a region of interest indicated by a solid line in FIG. 6 may be used.
  • a plurality of regions R1, R2, R3, R4, and R5 are provided in the second image, and the number of regions of interest equal to or less than the maximum number in each region R1, R2, R3, R4, and R5
  • a region of interest shown by a solid line may be used for setting of imaging conditions.
  • FIG. 7 shows the case where the maximum number is 3 in each of the regions R1, R2, R3, R4, and R5.
  • the operator may manually specify the region of interest used for setting the imaging conditions.
  • the moving mechanism 5 includes an X-axis ball screw 11 and an X-axis drive motor 12.
  • the first imaging unit 3 is fixed to the X-axis ball screw 11, and is moved in one direction (X direction) by rotating the X-axis ball screw 11 by the X-axis drive motor 12.
  • the second imaging unit 4 is fixed to the first imaging unit 3.
  • step S30 it is determined whether or not the distribution of change in exposure time is only one direction (direction of sub scanning) (step S30), and the distribution of change in exposure time is Whether or not to adjust the exposure time is determined regardless of whether it is only one direction (steps S20 and S31). If the distribution of the change in exposure time is only one direction and the exposure time is adjusted, it is set that the image is taken with the optimal exposure time for each line (step S32), and the process is ended.
  • step S33 when the distribution of the change in the exposure time is only one direction and the exposure time is not adjusted, it is set to capture an illumination light quantity corresponding to the optimal exposure time for each line (step S33), Finish. If the distribution of change in exposure time is not only one direction, and the exposure time is not adjusted, the process from step S20 is performed, and the process is ended.
  • the first imaging unit 3 and the second imaging unit 4 are moved in one direction (X direction) by the moving mechanism 5, and are line sensors in the direction (Y direction) orthogonal to the moving direction (X direction) by the moving mechanism 5. Since the first imaging unit 3 is moved, the first imaging unit 3 and the second imaging unit 4 can horizontally move in two directions (XY directions) orthogonal to each other.
  • an exchange mechanism that exchanges the optical system (first optical lens) of the first imaging unit 3 and the optical system (second optical lens) of the second imaging unit 4. May be employed.
  • the first imaging unit 3 and the second imaging unit 4 use a combination of an optical system and an imaging device for capturing light condensed by the optical system.
  • the first imaging unit 3 or the second imaging unit 4 is configured by exchanging the optical system by the exchange mechanism.
  • the cell image acquisition apparatus 1 includes a zoom optical system and an image pickup element for photographing light condensed by the zoom optical system, and the first image pickup unit according to the arrangement position of the zoom optical system.
  • the first imaging unit 3 is configured, and the zoom optical system is set to the second zoom position.
  • the second imaging unit 4 is configured by being disposed in

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Abstract

In order to acquire, in a short period of time, a high-contrast image covering a wide range of area of a culture vessel and properly manage the quality of a biological sample, this cell image acquisition device (1) is provided with: a first imaging unit (3) for imaging light from a cell (X) inside the culture vessel (2); a second imaging unit (4) for imaging a narrower visual field range at a higher resolution than with the first imaging unit; an area-of-interest setting unit (6) for setting one or more areas of interest that are narrower than a first visual field range which has been determined, as a result of analysis of a first image of the first visual field range acquired by the first imaging unit, to have a cell therein; and an imaging condition setting unit (7) for setting conditions for imaging to be performed by the first imaging unit on the basis of second images acquired, by the second imaging unit, of the respective areas of interest set by the area-of-interest setting unit, wherein a third image is acquired by the first imaging unit on the basis of the imaging conditions set by the imaging condition setting unit.

Description

細胞画像取得装置および細胞画像取得方法Cell image acquisition apparatus and cell image acquisition method
　本発明は、細胞画像取得装置および細胞画像取得方法に関するものである。 The present invention relates to a cell image acquisition apparatus and a cell image acquisition method.
　低倍率の撮影により取得された生体試料の広範囲な領域に対応する画像データにおいて観察対象を特定し、特定された観察対象について高倍率の撮影により高倍画像を取得し、取得された高倍画像をタイリングして、生体試料の広範囲な高倍率の画像を構築する観察装置が知られている（例えば、特許文献１参照。）。 An observation target is identified in image data corresponding to a wide area of a biological sample acquired by low magnification imaging, a high magnification image is acquired by high magnification imaging for the identified observation target, and the acquired high magnification image is tied An observation device is known which rings and constructs a wide range of high magnification images of a biological sample (see, for example, Patent Document 1).
特許第５５１６１０８号公報Patent No. 5516108 gazette
　しかしながら、高倍率の対物レンズの被写界深度は一般に極めて小さいため、生体試料の厚さ寸法が大きい場合には、焦点位置をずらしながら多数の画像を取得する必要があり、広範囲な領域についての画像を取得し終えるまでに多大な時間を要するという不都合がある。
　本発明は、上述した事情に鑑みてなされたものであって、培養容器の広範囲にわたる領域のコントラストの高い画像を短時間で取得して、生体試料の品質を適正に管理することができる細胞画像取得装置および細胞画像取得方法を提供することを目的としている。 However, since the depth of field of a high magnification objective lens is generally extremely small, when the thickness dimension of a biological sample is large, it is necessary to acquire a large number of images while shifting the focus position. There is a disadvantage that it takes a lot of time to finish acquiring an image.
The present invention has been made in view of the above-described circumstances, and it is possible to obtain a high-contrast image of a wide area of a culture vessel in a short time and to appropriately manage the quality of a biological sample It aims at providing an acquisition device and a cell image acquisition method.
　本発明の一態様は、培養容器内の細胞からの光を撮影する第１撮像部と、該第１撮像部よりも視野範囲が狭くかつ高解像度の撮影を行う第２撮像部と、前記第１撮像部により取得された第１画像を解析して細胞が存在すると判定された、前記第１撮像部の視野範囲より狭い１以上の関心領域を設定する関心領域設定部と、該関心領域設定部により設定された各前記関心領域について前記第２撮像部により取得された第２画像に基づいて、前記第１撮像部による撮影条件を設定する撮影条件設定部とを備え、該撮影条件設定部により設定された前記撮影条件に基づいて前記第１撮像部により第３画像を取得する細胞画像取得装置である。 One aspect of the present invention is a first imaging unit for imaging light from cells in a culture vessel, a second imaging unit for imaging with high resolution and a narrower visual field range than the first imaging unit, and 1. A region-of-interest setting unit for setting one or more regions of interest narrower than the field of view range of the first imaging unit determined by analyzing the first image acquired by the imaging unit and determining that there is a cell; An imaging condition setting unit configured to set an imaging condition by the first imaging unit based on a second image acquired by the second imaging unit for each of the regions of interest set by the unit; The first imaging unit is configured to acquire a third image based on the imaging condition set by the user.
　本態様によれば、培養容器内の細胞からの光を第１撮像部によって撮影することにより培養容器の第１画像が取得されると、関心領域設定部により第１画像が解析されて細胞が存在すると判定された１以上の関心領域が設定される。そして、第２撮像部により各関心領域について第２画像が取得される。 According to this aspect, when the first image of the culture container is acquired by photographing the light from the cells in the culture container by the first imaging unit, the first image is analyzed by the region of interest setting unit, and the cells are One or more regions of interest determined to exist are set. Then, a second image is acquired for each region of interest by the second imaging unit.
　第２画像は、第１撮像部による視野範囲より狭くかつ高解像度の撮影を行う第２撮像部により取得されているので、合焦範囲が狭く、かつ、コントラスト値の評価を厳密に行うことができる。したがって、撮影条件設定部により、第１撮像部による撮影条件を適正に設定することができ、設定された撮影条件に基づいて第１撮像部により第３画像を取得することにより、培養容器の広範囲にわたる領域のコントラストの高い画像を短時間で取得して、細胞の品質を適正に管理することができる。 Since the second image is acquired by the second imaging unit that performs imaging with high resolution and narrower than the visual field range by the first imaging unit, the focusing range is narrow and the contrast value is strictly evaluated. it can. Therefore, the imaging condition setting unit can appropriately set the imaging condition by the first imaging unit, and the third imaging unit acquires the third image by the first imaging unit based on the set imaging condition. The high-contrast image of the region can be acquired in a short time to properly manage the cell quality.
　上記態様においては、前記撮影条件設定部が、前記第１撮像部による合焦位置を設定してもよい。
　このようにすることで、第１撮像部による視野範囲より狭くかつ高解像度の撮影を行う第２撮像部により取得されることにより、合焦範囲が狭く、かつ、コントラスト値の評価を厳密に行うことができる画像を用いて、第１撮像部による適正な合焦位置を設定することができる。これにより、培養容器の広範囲にわたる領域のコントラストの高い画像を短時間で取得して、細胞の品質を適正に管理することができる。 In the above aspect, the photographing condition setting unit may set the in-focus position by the first imaging unit.
In this way, the focusing range is narrow and the contrast value is strictly evaluated by acquiring by the second imaging unit that performs imaging with high resolution and narrower than the visual field range by the first imaging unit. An appropriate in-focus position by the first imaging unit can be set using an image that can be captured. Thereby, high-contrast images of a wide area of the culture vessel can be obtained in a short time, and the cell quality can be properly managed.
　また、上記態様においては、前記撮影条件設定部が、前記第１撮像部による露光条件を設定してもよい。
　このようにすることで、第１撮像部による視野範囲より狭くかつ高解像度の撮影を行う第２撮像部により取得されることにより、合焦範囲が狭く、かつ、コントラスト値の評価を厳密に行うことができる画像を用いて、第１撮像部による適正な露光条件を設定することができる。これにより、培養容器の広範囲にわたる領域のコントラストの高い画像を短時間で取得して、細胞の品質を適正に管理することができる。 In the above aspect, the photographing condition setting unit may set an exposure condition by the first imaging unit.
In this way, the focusing range is narrow and the contrast value is strictly evaluated by acquiring by the second imaging unit that performs imaging with high resolution and narrower than the visual field range by the first imaging unit. An appropriate exposure condition by the first imaging unit can be set using an image that can be set. Thereby, high-contrast images of a wide area of the culture vessel can be obtained in a short time, and the cell quality can be properly managed.
　また、上記態様においては、前記関心領域設定部が、前記第１撮像部により取得された前記第１画像を複数の分割領域に分割し、各該分割領域において予め設定された数または予め設定された最大数以下の数の前記関心領域を設定してもよい。
　このようにすることで、関心領域として設定されるべき領域が多数存在する場合には、全ての領域について第２画像を取得するのではなく、予め制限された数の関心領域を設定することにより、処理を簡易にして処理時間を短縮することができる。 Further, in the above aspect, the region of interest setting unit divides the first image acquired by the first imaging unit into a plurality of divided regions, and the number or number set in advance is set in each of the divided regions. The number of regions of interest may be set equal to or less than the maximum number.
By doing this, when there are a large number of regions to be set as the regions of interest, it is possible to set a predetermined number of regions of interest instead of acquiring the second image for all the regions. The processing can be simplified and the processing time can be shortened.
　また、上記態様においては、第１光学レンズおよび第２光学レンズと、前記第１光学レンズと前記第２光学レンズとを交換する交換機構と、前記第１光学レンズまたは前記第２光学レンズにより集光された光を撮影する撮像素子とを備え、前記第１撮像部が、前記第１光学レンズと前記撮像素子との組合せにより構成され、前記第２撮像部が、前記第２光学レンズと前記撮像素子との組合せにより構成されていてもよい。 In the above aspect, the first optical lens and the second optical lens, the exchange mechanism for exchanging the first optical lens and the second optical lens, and the collection by the first optical lens or the second optical lens And an imaging device configured to capture the emitted light, wherein the first imaging unit is configured by a combination of the first optical lens and the imaging device, and the second imaging unit is the second optical lens and the second imaging unit. You may be comprised by the combination with an image pick-up element.
　また、上記態様においては、ズーム光学系と、該ズーム光学系により集光された光を撮影する撮像素子とを備え、前記第１撮像部が、前記ズーム光学系を第１ズーム位置に配置することにより構成され、前記第２撮像部が、前記ズーム光学系を第２ズーム位置に配置することにより構成されていてもよい。 Further, in the above aspect, the zoom optical system includes the zoom optical system and an image pickup element for photographing light condensed by the zoom optical system, and the first image pickup unit arranges the zoom optical system at a first zoom position. The second imaging unit may be configured by disposing the zoom optical system at a second zoom position.
　また、本発明の他の態様は、培養容器内の細胞からの光を撮影して第１視野範囲の第１画像を取得する第１撮影ステップと、該第１撮影ステップにおいて取得された前記第１画像を解析して前記細胞が存在すると判定された前記第１視野範囲より狭い１以上の関心領域を設定する関心領域設定ステップと、該関心領域設定ステップにおいて設定された各前記関心領域について前記第１撮影ステップよりも高解像度の撮影を行って１以上の第２画像を取得する第２撮影ステップと、該第２撮影ステップにおいて取得された１以上の前記第２画像に基づいて、撮影条件を設定する撮影条件設定ステップと、該撮影条件設定ステップにおいて設定された前記撮影条件に基づいて前記第１視野範囲について前記第１撮影ステップと同じ解像度の撮影を行う第３撮影ステップとを含む細胞画像取得方法である。 Moreover, the other aspect of this invention is a 1st imaging | photography step which image | photographs the light from the cell in a culture container, and acquires the 1st image of the 1st visual field range, Said said imaging | photography step acquired A region of interest setting step of analyzing one image to set one or more regions of interest narrower than the first visual field range determined to have the cells present, and the respective regions of interest set in the region of interest setting step A shooting condition based on a second shooting step of shooting at a resolution higher than that of the first shooting step to obtain one or more second images, and the one or more second images obtained in the second shooting step A shooting condition setting step of setting the shooting condition, and shooting of the same resolution as the first shooting step for the first visual field range based on the shooting condition set in the shooting condition setting step A cell image acquiring method and a third imaging step of performing.
　本発明によれば、培養容器の広範囲にわたる領域のコントラストの高い画像を短時間で取得して、生体試料の品質を適正に管理することができるという効果を奏する。 ADVANTAGE OF THE INVENTION According to this invention, it is effective in the ability to acquire the high contrast image of the wide area | region of a culture container in a short time, and to manage the quality of a biological sample appropriately.
本発明の一実施形態に係る細胞画像取得装置を示す全体構成図である。BRIEF DESCRIPTION OF THE DRAWINGS It is a whole block diagram which shows the cell image acquisition apparatus which concerns on one Embodiment of this invention. 図１の細胞画像取得装置のベース部を示す縦断面図である。It is a longitudinal cross-sectional view which shows the base part of the cell image acquisition apparatus of FIG. 図１の細胞画像取得装置のベース部を示す上面図である。It is a top view which shows the base part of the cell image acquisition apparatus of FIG. 図１の細胞画像取得装置における露出条件の設定方法を説明するフローチャートである。It is a flowchart explaining the setting method of the exposure conditions in the cell image acquisition apparatus of FIG. 図１の細胞画像取得装置を用いた細胞画像取得方法を説明するフローチャートである。It is a flowchart explaining the cell image acquisition method using the cell image acquisition apparatus of FIG. 図１の細胞画像取得装置により取得された第１画像内から一部の関心領域を撮影条件設定用に設定する一例を示す図である。It is a figure which shows an example which sets a part of interest area | region from the inside of the 1st image acquired by the cell image acquisition apparatus of FIG. 1 for imaging condition setting. 図１の細胞画像取得装置により取得された第２画像内から一部の関心領域を撮影条件設定用に設定する他の例を示す図である。It is a figure which shows the other example which sets a part of interest area | region from the inside of the 2nd image acquired by the cell image acquisition apparatus of FIG. 1 for imaging condition setting. 図１の細胞画像取得装置の変形例のベース部を示す縦断面図である。It is a longitudinal cross-sectional view which shows the base part of the modification of the cell image acquisition apparatus of FIG. 図１の細胞画像取得装置の変形例のベース部を示す上面図である。It is a top view which shows the base part of the modification of the cell image acquisition apparatus of FIG. 図１の細胞画像取得装置の変形例における露出条件の設定方法を説明するフローチャートである。It is a flowchart explaining the setting method of the exposure conditions in the modification of the cell image acquisition apparatus of FIG.
　本発明の一実施形態に係る細胞画像取得装置１および細胞画像取得方法について、図面を参照して以下に説明する。
　本実施形態に係る細胞画像取得装置１は、培養容器２内に収容されて培養されている細胞Ｘの画像を取得する装置であって、図１から図３に示されるように、視野範囲および解像度の異なる第１撮像部３および第２撮像部４と、これらの撮像部３，４を水平移動させる移動機構５とを備えている。第１撮像部３、第２撮像部４および移動機構５によりベース部１０が構成されている。 A cell image acquisition apparatus 1 and a cell image acquisition method according to an embodiment of the present invention will be described below with reference to the drawings.
The cell image acquisition apparatus 1 according to the present embodiment is an apparatus for acquiring an image of the cell X accommodated and cultured in the culture container 2, and as shown in FIGS. 1 to 3, the visual field range and The first imaging unit 3 and the second imaging unit 4 having different resolutions, and the moving mechanism 5 for horizontally moving the imaging units 3 and 4 are provided. The base unit 10 is configured by the first imaging unit 3, the second imaging unit 4, and the moving mechanism 5.
　第１撮像部３は、例えば、移動機構５により一方向に移動させられるとともに、移動機構５による移動方向に交差する方向に配列された複数の画素を備えるラインセンサであり、移動機構５によって移動させられる間に、ライン状の画像を順次取得していくことにより、培養容器２の底面の略全面にわたる広範囲な第１視野範囲の第１画像を取得するようになっている。なお、第１視野範囲は必ずしも培養容器２の底面の略全面にわたっていなくてもよい。
　また、第１撮像部３としては、ラインセンサに限定されるものではなく、他の任意の撮像部を採用してもよい。 The first imaging unit 3 is, for example, a line sensor including a plurality of pixels which are moved in one direction by the moving mechanism 5 and arranged in a direction crossing the moving direction by the moving mechanism 5. By sequentially acquiring line-shaped images while being driven, a first image of a wide first field of view over substantially the entire bottom surface of the culture vessel 2 is acquired. Note that the first visual field range may not necessarily cover substantially the entire bottom surface of the culture vessel 2.
Further, the first imaging unit 3 is not limited to the line sensor, and any other imaging unit may be adopted.
　第２撮像部４は、例えば、第１撮像部３よりも狭い第２視野範囲の第１撮像部３より高解像度の第２画像を取得する図示しない固体撮像素子を備える。移動機構５の作動により第２撮像部４を２次元的に水平方向に移動させることにより、移動先の各位置において第２画像を取得することができる。 The second imaging unit 4 includes, for example, a solid-state imaging device (not shown) that acquires a second image of high resolution from the first imaging unit 3 in a second visual field range narrower than the first imaging unit 3. By moving the second imaging unit 4 in a two-dimensional horizontal direction by the operation of the moving mechanism 5, it is possible to acquire a second image at each position of the movement destination.
　第１撮像部３および第２撮像部４は図示しない合焦機構および露出調整機構を備えている。
　合焦機構は、例えば、１以上のレンズを光軸方向に移動させる機構である。また、露出調整機構は、例えば、シャッタである。 The first imaging unit 3 and the second imaging unit 4 are provided with a focusing mechanism and an exposure adjustment mechanism not shown.
The focusing mechanism is, for example, a mechanism that moves one or more lenses in the optical axis direction. The exposure adjustment mechanism is, for example, a shutter.
　移動機構５は、図２および図３に示されるように、Ｘ軸ボールねじ１１と、該Ｘ軸ボールねじ１１を回転させるＸ軸駆動モータ１２と、該Ｘ軸駆動モータ１２によって回転させられたＸ軸ボールねじ１１上を一方向（Ｘ方向）に移動するＹ軸駆動モータ１３と、Ｘ軸ボールねじ１１と直交するように配置され、Ｙ軸駆動モータ１３により回転させられるＹ軸ボールねじ１４と、Ｙ軸駆動モータ１３によって回転させられたＹ軸ボールねじ１４上を一方向（Ｙ方向）に移動するスライダ１５とを備えている。
　スライダ１５は、第１撮像部３および第２撮像部４が固定されており、相互に直交する２方向（ＸＹ方向）に水平移動するようになっている。 The moving mechanism 5 is rotated by an X-axis drive motor 12 that rotates the X-axis ball screw 11, the X-axis ball screw 11, and the X-axis drive motor 12 as shown in FIGS. 2 and 3. A Y-axis drive screw 13 which moves on the X-axis ball screw 11 in one direction (X-direction), and a Y-axis ball screw 14 which is disposed orthogonal to the X-axis ball screw 11 and is rotated by the Y-axis drive motor 13 And a slider 15 moving in one direction (Y direction) on the Y-axis ball screw 14 rotated by the Y-axis drive motor 13.
The first imaging unit 3 and the second imaging unit 4 are fixed to the slider 15, and the slider 15 horizontally moves in two directions (XY directions) orthogonal to each other.
　また、本実施形態に係る細胞画像取得装置１は、図１に示されるように、第１撮像部３に接続された関心領域設定部６と、第２撮像部４に接続された撮影条件設定部７と、第１撮像部３、第２撮像部４および移動機構５を制御する制御部８と、最終的に第１撮像部３により取得された第３画像を表示する表示部９とを備えている。なお、表示部９に代えてあるいは表示部９に加えて、第３画像を記憶する記憶部を備えていてもよい。 In addition, as shown in FIG. 1, the cell image acquisition apparatus 1 according to the present embodiment has the region of interest setting unit 6 connected to the first imaging unit 3 and the imaging condition setting connected to the second imaging unit 4. A control unit 8 for controlling the unit 7, the first imaging unit 3, the second imaging unit 4 and the moving mechanism 5, and a display unit 9 for displaying the third image finally acquired by the first imaging unit 3; Have. A storage unit for storing the third image may be provided instead of or in addition to the display unit 9.
　関心領域設定部６は、第１撮像部３により取得された広範囲な第１視野範囲にわたる第１画像を解析して、細胞Ｘが存在するか否かを判定し、細胞Ｘが存在すると判定された領域を、第１視野範囲より十分に小さい関心領域（ＲＯＩ）として設定するようになっている。具体的には、設定した関心領域の中心座標を記憶するようになっている。
　細胞Ｘが存在すると判定された領域が複数存在する場合には、複数の関心領域が設定され、関心領域よりも広い範囲にわたって細胞Ｘが存在すると判定された領域が存在する場合には、複数の関心領域が隣接して設定されるようになる。 The region of interest setting unit 6 analyzes the first image over the wide first field range acquired by the first imaging unit 3 to determine whether or not the cell X is present, and it is determined that the cell X is present. The region of interest is set as a region of interest (ROI) sufficiently smaller than the first visual field range. Specifically, the center coordinates of the set region of interest are stored.
If there are a plurality of regions determined that the cell X is present, a plurality of regions of interest are set, and if there are regions determined that the cell X is present over a wider range than the regions of interest, a plurality of regions are determined. Regions of interest will be set adjacent to each other.
　撮影条件設定部７は、第２撮像部により取得された関心領域毎の第２画像を用いて、各関心領域における最適な合焦位置および最適な露出条件（露光条件）を求めるようになっている。具体的には、合焦位置については、各関心領域において第２撮像部４による合焦位置を調整しながら撮影を行い、合焦範囲を検出するようになっている。そして、検出された合焦範囲が、第１撮像部３の被写界深度を超えていない場合には、合焦範囲の中央位置を合焦位置とし、合焦範囲が第１撮像部３の被写界深度を超えている場合には、合焦範囲を被写界深度相当で分割し各被写界深度の中央位置に合焦位置を設定するようになっている。 The imaging condition setting unit 7 obtains the optimal focusing position and the optimal exposure condition (exposure condition) in each region of interest, using the second image for each region of interest acquired by the second imaging unit. There is. Specifically, with regard to the in-focus position, imaging is performed while adjusting the in-focus position by the second imaging unit 4 in each region of interest, and the in-focus range is detected. When the detected in-focus range does not exceed the depth of field of the first imaging unit 3, the center position of the in-focus range is set as the in-focus position, and the in-focus range is the first imaging unit 3. When the depth of field is exceeded, the focusing range is divided by the depth of field and the focusing position is set at the center position of each depth of field.
　また、露出条件については、図４に示されるように、まず、露光時間を調整するか否かが判定され（ステップＳ２０）、露光時間の調整の有無に係わらず、画像の飽和を許すか否かが判定される（ステップＳ２１，Ｓ２２）。
　露光時間を調整し、かつ画像の飽和を許す場合には平均の露光時間で撮影するように設定され（ステップＳ２３）、処理を終了する。 As for the exposure condition, as shown in FIG. 4, first, it is judged whether or not the exposure time is adjusted (step S20), and whether or not the image is allowed to be saturated regardless of the adjustment of the exposure time Is determined (steps S21 and S22).
The exposure time is adjusted, and when saturation of the image is allowed, it is set to shoot at the average exposure time (step S23), and the process is ended.
　また、露光時間を調整し、かつ画像の飽和を許さない場合には最小から最大までの露光時間で少なくとも２回以上撮影するように設定され（ステップＳ２４）、処理を終了する。
　また、露光時間を調整せず、かつ画像の飽和を許す場合には平均の露光時間に相当する照明光量で撮影するように設定され（ステップＳ２５）、処理を終了する。 Further, the exposure time is adjusted, and when the saturation of the image is not permitted, it is set to shoot at least twice or more with the exposure time from the minimum to the maximum (step S24), and the process is ended.
If the exposure time is not adjusted and saturation of the image is allowed, shooting is performed with the illumination light amount corresponding to the average exposure time (step S25), and the process is ended.
　また、露光時間を調整せず、かつ画像の飽和を許さない場合には最小から最大までの露光時間に相当する照明光量で撮影するように設定され（ステップＳ２６）、処理を終了する。例えば、第２撮像部４が備えている自動露出機能を利用して、各関心領域において適正なシャッタスピードを検出するようになっている。露出条件としては、シャッタスピード、図示しない照明光源から射出される照明光の光量、あるいは絞りの開度等によって調整することができる。
　シャッタスピードは、第１撮像部３の画素配列方向には検出されたシャッタスピードの平均値を設定し、移動機構５による移動方向にはシャッタスピードを変化させながら第１撮像部３による撮影を行うように設定される。 If the exposure time is not adjusted and saturation of the image is not permitted, shooting is performed with the illumination light amount corresponding to the minimum to maximum exposure time (step S26), and the process is ended. For example, an appropriate shutter speed is detected in each region of interest using the automatic exposure function of the second imaging unit 4. The exposure condition can be adjusted by the shutter speed, the light amount of the illumination light emitted from the illumination light source (not shown), the opening degree of the aperture, or the like.
The shutter speed is set to the average value of the detected shutter speeds in the pixel array direction of the first imaging unit 3 and the first imaging unit 3 performs imaging while changing the shutter speed in the moving direction by the moving mechanism 5 Is set as
　このように構成された本実施形態に係る細胞画像取得装置１を用いた細胞画像取得方法について、以下に説明する。
　本実施形態に係る細胞画像取得装置１を用いて、培養容器２の底面の画像を取得するには、図５に示されるように、まず、制御部８は、第１撮像部３の合焦位置を培養容器２の底面より若干上方に設定して、移動機構５により第１撮像部３を移動させながら培養容器２の底面のほぼ全面にわたる第１視野範囲の第１画像を取得（第１撮影）させる（第１撮影ステップＳ１）。 A cell image acquisition method using the cell image acquisition device 1 according to the present embodiment configured as described above will be described below.
In order to acquire an image of the bottom of the culture container 2 using the cell image acquisition device 1 according to the present embodiment, as shown in FIG. 5, first, the control unit 8 controls the focus of the first imaging unit 3. The first image of the first field of view over the entire bottom surface of the culture container 2 is obtained while setting the position slightly above the bottom surface of the culture container 2 and moving the first imaging unit 3 by the moving mechanism 5 (first Shooting) (first shooting step S1).
　第１撮像部３により第１画像が取得されると、取得された第１画像は関心領域設定部６に送られて、細胞Ｘが存在するか否かの解析が行われ、細胞Ｘが存在すると認められた領域が関心領域として設定される（関心領域設定ステップＳ２）。設定された各関心領域の座標（例えば、中心座標）は記憶される（ステップＳ３）。 When the first image is acquired by the first imaging unit 3, the acquired first image is sent to the region of interest setting unit 6, and analysis of whether or not the cell X is present is performed, and the cell X is present. Then, the recognized region is set as a region of interest (region of interest setting step S2). The coordinates (for example, center coordinates) of each set region of interest are stored (step S3).
　全ての関心領域の座標が記憶された後には、制御部８は、移動機構５により第２撮像部４を移動させ、第２撮像部４の第２視野範囲をいずれかの関心領域に一致させる（ステップＳ４）。この状態で、制御部８は第２撮像部４の合焦位置を光軸方向に変化させながら第２撮像部４により第２画像を取得（第２撮影）させる（第２撮影ステップＳ５）。 After the coordinates of all the regions of interest are stored, the control unit 8 causes the moving mechanism 5 to move the second imaging unit 4 so that the second view range of the second imaging unit 4 matches any of the regions of interest. (Step S4). In this state, the control unit 8 causes the second imaging unit 4 to acquire a second image (second imaging) while changing the in-focus position of the second imaging unit 4 in the optical axis direction (second imaging step S5).
　第２撮像部４により第２画像が取得されると、撮影条件設定部７が、光軸方向に細胞Ｘが存在する範囲を合焦範囲として求めるとともに、該合焦範囲において適正露出となるシャッタスピードを露出条件として検出し記憶する（撮影条件設定ステップＳ６）。 When the second image is acquired by the second imaging unit 4, the imaging condition setting unit 7 determines a range in which the cell X exists in the optical axis direction as a focusing range, and a shutter that provides appropriate exposure in the focusing range The speed is detected as an exposure condition and stored (shooting condition setting step S6).
　全ての関心領域について合焦範囲および露出条件が検出されたか否かが判定され（ステップＳ７）、終了していない場合には、ステップＳ４からの工程が繰り返される。全ての関心領域について合焦範囲および露出条件が検出された場合には、撮影条件設定部７は、全ての関心領域の合焦範囲から培養容器２全体の合焦範囲を求める（ステップＳ８）。 It is determined whether the in-focus range and the exposure condition have been detected for all the regions of interest (step S7), and if not completed, the process from step S4 is repeated. When the in-focus range and the exposure condition are detected for all the regions of interest, the imaging condition setting unit 7 obtains the in-focus range of the entire culture vessel 2 from the in-focus regions of all the areas of interest (step S8).
　そして、撮影条件設定部７においては、培養容器２全体の合焦範囲が第１撮像部３による被写界深度を超えているか否かが判定され（ステップＳ９）、超えている場合には、合焦範囲を第１撮像部３による被写界深度によって複数ブロックに分割し、ブロック毎に合焦位置および露出条件を設定する（ステップＳ１０）。超えていない場合には、後述するステップＳ１１の工程を実行する。
　また、第１撮像部３の画素配列方向に複数の関心領域が存在する場合には、移動機構５による移動方向に異なる撮影位置毎に、複数の関心領域について算出されたシャッタスピードの平均値がシャッタスピードとして設定される。 Then, in the imaging condition setting unit 7, it is determined whether the in-focus range of the entire culture container 2 exceeds the depth of field by the first imaging unit 3 (step S9). The focusing range is divided into a plurality of blocks according to the depth of field by the first imaging unit 3, and the focusing position and the exposure condition are set for each block (step S10). If not exceeded, the process of step S11 described later is executed.
In addition, when a plurality of regions of interest exist in the pixel array direction of the first imaging unit 3, the average value of the shutter speeds calculated for the plurality of regions of interest is different for each imaging position different in the movement direction by the moving mechanism 5. The shutter speed is set.
　その後、制御部８は、撮影条件設定部７により設定された撮影条件を用いて、再度、第１撮像部３を作動させ、第１撮像部３により第１視野範囲の第３画像を取得（第３撮影）させる（第３撮影ステップＳ１１）。
　第３撮影ステップＳ１１においては、ステップＳ１０において、複数ブロックの撮影条件が設定されている場合には、ブロック毎の合焦位置および露出条件に従って、ブロック数に等しい回数だけ第３画像の取得が行われる。 After that, the control unit 8 causes the first imaging unit 3 to operate again using the imaging conditions set by the imaging condition setting unit 7, and the first imaging unit 3 acquires the third image of the first visual field range ( The third shooting) is performed (third shooting step S11).
In the third imaging step S11, when imaging conditions of a plurality of blocks are set in step S10, acquisition of the third image is performed a number of times equal to the number of blocks according to the in-focus position and exposure condition for each block. It will be.
　このように、本実施形態に係る細胞画像取得装置１および細胞画像取得方法によれば、広範囲の第１視野範囲を有する第１撮像部３によって取得された広範囲の第１画像に基づいて関心領域を検出し、各関心領域について、第１撮像部３より視野が狭く高解像の第２撮像部４によって取得された第２画像に基づいて撮影条件を設定する。その結果、合焦範囲が狭く、かつ、コントラスト値の評価を厳密に行うことができる第２画像によって、第１撮像部３による撮影条件を適正に設定することができる。 As described above, according to the cell image acquisition device 1 and the cell image acquisition method according to the present embodiment, the region of interest is based on the wide-area first image acquired by the first imaging unit 3 having the wide-range first visual field range. Are detected, and imaging conditions are set based on the second image acquired by the second imaging unit 4 having a narrower field of view than the first imaging unit 3 and high resolution for each region of interest. As a result, the imaging condition by the first imaging unit 3 can be appropriately set by the second image in which the focusing range is narrow and the evaluation of the contrast value can be strictly performed.
　そして、設定された撮影条件に基づいて第１撮像部３により第３画像を取得することにより、培養容器２の広範囲にわたる領域のコントラストの高い画像を短時間で取得することができ、細胞Ｘの品質管理を迅速に行うことができるという利点がある。 Then, by acquiring the third image by the first imaging unit 3 based on the set imaging conditions, it is possible to acquire an image with high contrast in a wide area of the culture vessel 2 in a short time. There is an advantage that quality control can be performed quickly.
　また、第２撮像部４により検出された合焦範囲が、第１撮像部３の被写界深度を超えている場合には、合焦範囲を分割して複数回に分けて第３画像を取得するので、細胞Ｘが成長して高さ方向の寸法が大きくなっても、細胞Ｘが存在する範囲を取りこぼしなく観察することができるという利点がある。例えば、培養容器２の底面が平面ではなく高低差がある場合等においても、細胞Ｘを取りこぼしなく観察することができる。 When the in-focus range detected by the second imaging unit 4 exceeds the depth of field of the first imaging unit 3, the in-focus range is divided and divided into a plurality of times to divide the third image. Since the cells X are obtained, even if the cell X grows and the size in the height direction is increased, there is an advantage that the range in which the cell X is present can be observed without being missed. For example, even in the case where the bottom surface of the culture container 2 is not flat but has a height difference, etc., it is possible to observe the cell X without losing it.
　なお、本実施形態においては、検出された全ての関心領域について第２画像を取得することとしたが、これに代えて、一部の関心領域のみを用いて撮影条件を設定することにしてもよい。
　例えば、図６に実線および破線で示されるように、複数の関心領域が検出された場合に、第１画像を複数の領域Ｒ１，Ｒ２，Ｒ３，Ｒ４に分割し、各領域Ｒ１，Ｒ２，Ｒ３，Ｒ４内に存在するいずれか１つの関心領域を撮影条件の設定のために使用することにしてもよい。例えば、図６に実線で示される関心領域を使用すればよい。 In the present embodiment, the second image is acquired for all the detected regions of interest, but instead of this, the imaging conditions may be set using only a part of the regions of interest Good.
For example, as shown by a solid line and a broken line in FIG. 6, when a plurality of regions of interest are detected, the first image is divided into a plurality of regions R1, R2, R3, R4, and each region R1, R2, R3 is , R4 may be used for setting the imaging conditions. For example, a region of interest indicated by a solid line in FIG. 6 may be used.
　また、図７に示されるように、第２画像内に複数の領域Ｒ１，Ｒ２，Ｒ３，Ｒ４，Ｒ５を設け、各領域Ｒ１，Ｒ２，Ｒ３，Ｒ４，Ｒ５において、最大数以下の関心領域（図７中、実線で示される関心領域）を撮影条件の設定のために使用することにしてもよい。図７は、各領域Ｒ１，Ｒ２，Ｒ３，Ｒ４，Ｒ５内において最大数３とした場合を示している。
　また、撮影条件の設定に使用する関心領域を作業者が手動で指定することにしてもよい。 Further, as shown in FIG. 7, a plurality of regions R1, R2, R3, R4, and R5 are provided in the second image, and the number of regions of interest equal to or less than the maximum number in each region R1, R2, R3, R4, and R5 In FIG. 7, a region of interest shown by a solid line may be used for setting of imaging conditions. FIG. 7 shows the case where the maximum number is 3 in each of the regions R1, R2, R3, R4, and R5.
In addition, the operator may manually specify the region of interest used for setting the imaging conditions.
　また、本実施形態においては、第１撮像部３として、エリアセンサを採用したものを例示したが、これに代えて、ラインセンサを採用してもよい。
　この場合、図８および図９に示されるように、移動機構５は、Ｘ軸ボールねじ１１と、Ｘ軸駆動モータ１２とを備えている。
　第１撮像部３は、Ｘ軸ボールねじ１１に固定されており、Ｘ軸駆動モータ１２によってＸ軸ボールねじ１１が回転させられることにより一方向（Ｘ方向）に移動するようになっている。また、第２撮像部４は、第１撮像部３に固定されている。 Moreover, although what employ | adopted the area sensor as the 1st imaging part 3 was illustrated in this embodiment, it may replace with this and may employ | adopt a line sensor.
In this case, as shown in FIGS. 8 and 9, the moving mechanism 5 includes an X-axis ball screw 11 and an X-axis drive motor 12.
The first imaging unit 3 is fixed to the X-axis ball screw 11, and is moved in one direction (X direction) by rotating the X-axis ball screw 11 by the X-axis drive motor 12. The second imaging unit 4 is fixed to the first imaging unit 3.
　露出条件については、図１０に示されるように、まず、露光時間の変化の分布が一方向（副走査の方向）のみであるか否か判定され（ステップＳ３０）、露光時間の変化の分布が一方向のみであるか否かに係わらず、露光時間を調整するか否かが判定される（ステップＳ２０，Ｓ３１）。
　露光時間の変化の分布が一方向のみであり、かつ露光時間を調整する場合には、ラインごとに最適な露光時間で撮影するように設定され（ステップＳ３２）、処理を終了する。 As for the exposure condition, as shown in FIG. 10, first, it is determined whether or not the distribution of change in exposure time is only one direction (direction of sub scanning) (step S30), and the distribution of change in exposure time is Whether or not to adjust the exposure time is determined regardless of whether it is only one direction (steps S20 and S31).
If the distribution of the change in exposure time is only one direction and the exposure time is adjusted, it is set that the image is taken with the optimal exposure time for each line (step S32), and the process is ended.
　また、露光時間の変化の分布が一方向のみであり、かつ露光時間を調整しない場合には、ラインごとに最適な露光時間に相当する照明光量で撮影するように設定され（ステップＳ３３）、処理を終了する。
　また、露光時間の変化の分布が一方向のみではなく、かつ露光時間を調整しない場合には、ステップＳ２０からの工程を実行し、処理を終了する。 Further, when the distribution of the change in the exposure time is only one direction and the exposure time is not adjusted, it is set to capture an illumination light quantity corresponding to the optimal exposure time for each line (step S33), Finish.
If the distribution of change in exposure time is not only one direction, and the exposure time is not adjusted, the process from step S20 is performed, and the process is ended.
　第１撮像部３および第２撮像部４は、移動機構５によって一方向（Ｘ方向）に移動され、移動機構５による移動方向（Ｘ方向）に直交する方向（Ｙ方向）においてはラインセンサである第１撮像部３により移動させられるため、第１撮像部３および第２撮像部４が相互に直交する２方向（ＸＹ方向）に水平移動することができる。 The first imaging unit 3 and the second imaging unit 4 are moved in one direction (X direction) by the moving mechanism 5, and are line sensors in the direction (Y direction) orthogonal to the moving direction (X direction) by the moving mechanism 5. Since the first imaging unit 3 is moved, the first imaging unit 3 and the second imaging unit 4 can horizontally move in two directions (XY directions) orthogonal to each other.
　また、本実施形態においては、細胞画像取得装置１として、第１撮像部３の光学系（第１光学レンズ）と第２撮像部４の光学系（第２光学レンズ）とを交換する交換機構を備えるものを採用してもよい。
　この場合、第１撮像部３および第２撮像部４は、光学系と該光学系により集光された光を撮影する撮像素子との組み合わせにより構成されるものを用いる。交換機構によって光学系を交換することにより、第１撮像部３または第２撮像部４が構成される。 Further, in the present embodiment, as the cell image acquisition device 1, an exchange mechanism that exchanges the optical system (first optical lens) of the first imaging unit 3 and the optical system (second optical lens) of the second imaging unit 4. May be employed.
In this case, the first imaging unit 3 and the second imaging unit 4 use a combination of an optical system and an imaging device for capturing light condensed by the optical system. The first imaging unit 3 or the second imaging unit 4 is configured by exchanging the optical system by the exchange mechanism.
　また、本実施形態においては、細胞画像取得装置１として、ズーム光学系と、該ズーム光学系により集光された光を撮影する撮像素子とを備え、ズーム光学系の配置位置によって第１撮像部３または第２撮像部４を構成するものを採用してもよい
　この場合、ズーム光学系を第１ズーム位置に配置することにより第１撮像部３が構成され、ズーム光学系を第２ズーム位置に配置することにより第２撮像部４が構成される。 Further, in the present embodiment, the cell image acquisition apparatus 1 includes a zoom optical system and an image pickup element for photographing light condensed by the zoom optical system, and the first image pickup unit according to the arrangement position of the zoom optical system. In this case, by arranging the zoom optical system at the first zoom position, the first imaging unit 3 is configured, and the zoom optical system is set to the second zoom position. The second imaging unit 4 is configured by being disposed in
　１　細胞画像取得装置
　２　培養容器
　３　第１撮像部
　４　第２撮像部
　６　関心領域設定部
　７　撮影条件設定部
　Ｘ　細胞
　Ｓ１　第１撮影ステップ
　Ｓ２　関心領域設定ステップ
　Ｓ５　第２撮影ステップ
　Ｓ６　撮影条件設定ステップ
　Ｓ１１　第３撮影ステップ DESCRIPTION OF SYMBOLS 1 cell image acquisition apparatus 2 culture container 3 1st imaging part 4 2nd imaging part 6 region of interest setting part 7 imaging condition setting part X cell S1 1st imaging step S2 area of interest setting step S5 2nd imaging step S6 imaging condition setting step S11 3rd shooting step

Claims (7)

  1. 　培養容器内の細胞からの光を撮影する第１撮像部と、
    　該第１撮像部よりも視野範囲が狭くかつ高解像度の撮影を行う第２撮像部と、
    　前記第１撮像部により取得された第１画像を解析して細胞が存在すると判定された、前記第１撮像部の視野範囲より狭い１以上の関心領域を設定する関心領域設定部と、
    　該関心領域設定部により設定された各前記関心領域について前記第２撮像部により取得された第２画像に基づいて、前記第１撮像部による撮影条件を設定する撮影条件設定部とを備え、
    　該撮影条件設定部により設定された前記撮影条件に基づいて前記第１撮像部により第３画像を取得する細胞画像取得装置。     A first imaging unit configured to capture light from cells in the culture vessel;
    A second imaging unit that performs high-resolution imaging with a narrower visual field range than the first imaging unit;
    A region of interest setting unit configured to set one or more regions of interest narrower than the field of view range of the first imaging unit, which is determined by analyzing the first image acquired by the first imaging unit and determining that there is a cell;
    An imaging condition setting unit configured to set an imaging condition by the first imaging unit based on a second image acquired by the second imaging unit for each of the regions of interest set by the region of interest setting unit;
    A cell image acquisition apparatus for acquiring a third image by the first imaging unit based on the imaging condition set by the imaging condition setting unit.
  2. 　前記撮影条件設定部が、前記第１撮像部による合焦位置を設定する請求項１に記載の細胞画像取得装置。 The cell image acquisition device according to claim 1, wherein the imaging condition setting unit sets an in-focus position by the first imaging unit.
  3. 　前記撮影条件設定部が、前記第１撮像部による露光条件を設定する請求項１または請求項２に記載の細胞画像取得装置。 The cell image acquisition apparatus according to claim 1, wherein the imaging condition setting unit sets an exposure condition by the first imaging unit.
  4. 　前記関心領域設定部が、前記第１撮像部により取得された前記第１画像を複数の分割領域に分割し、各該分割領域において予め設定された数または予め設定された最大数以下の数の前記関心領域を設定する請求項１から請求項３のいずれかに記載の細胞画像取得装置。 The region-of-interest setting unit divides the first image acquired by the first imaging unit into a plurality of divided regions, and the number set in advance in each divided region or a number smaller than the maximum number set in advance The cell image acquisition device according to any one of claims 1 to 3, wherein the region of interest is set.
  5. 　第１光学レンズおよび第２光学レンズと、前記第１光学レンズと前記第２光学レンズとを交換する交換機構と、前記第１光学レンズまたは前記第２光学レンズにより集光された光を撮影する撮像素子とを備え、
    　前記第１撮像部が、前記第１光学レンズと前記撮像素子との組合せにより構成され、
    　前記第２撮像部が、前記第２光学レンズと前記撮像素子との組合せにより構成される請求項１から請求項４のいずれかに記載の細胞画像取得装置。     The first optical lens and the second optical lens, the exchange mechanism for exchanging the first optical lens and the second optical lens, and the light collected by the first optical lens or the second optical lens And an imaging device,
    The first imaging unit is configured by a combination of the first optical lens and the imaging device.
    The cell image acquisition device according to any one of claims 1 to 4, wherein the second imaging unit is configured by a combination of the second optical lens and the imaging element.
  6. 　ズーム光学系と、該ズーム光学系により集光された光を撮影する撮像素子とを備え、
    　前記第１撮像部が、前記ズーム光学系を第１ズーム位置に配置することにより構成され、
    　前記第２撮像部が、前記ズーム光学系を第２ズーム位置に配置することにより構成される請求項１から請求項４のいずれかに記載の細胞画像取得装置。     A zoom optical system, and an image pickup device for photographing light condensed by the zoom optical system;
    The first imaging unit is configured by arranging the zoom optical system at a first zoom position,
    The cell image acquisition device according to any one of claims 1 to 4, wherein the second imaging unit is configured by arranging the zoom optical system at a second zoom position.
  7. 　培養容器内の細胞からの光を撮影して第１視野範囲の第１画像を取得する第１撮影ステップと、
    　該第１撮影ステップにおいて取得された前記第１画像を解析して前記細胞が存在すると判定された、前記第１視野範囲より狭い１以上の関心領域を設定する関心領域設定ステップと、
    　該関心領域設定ステップにおいて設定された各前記関心領域について前記第１撮影ステップよりも高解像度の撮影を行って１以上の第２画像を取得する第２撮影ステップと、
    　該第２撮影ステップにおいて取得された１以上の前記第２画像に基づいて、撮影条件を設定する撮影条件設定ステップと、
    　該撮影条件設定ステップにおいて設定された前記撮影条件に基づいて前記第１視野範囲について前記第１撮影ステップと同じ解像度の撮影を行う第３撮影ステップとを含む細胞画像取得方法。
          A first imaging step of imaging light from cells in the culture vessel to acquire a first image of a first visual field range;
    A region of interest setting step of setting one or more regions of interest narrower than the first visual field range, wherein the first image acquired in the first imaging step is analyzed to determine that the cell is present;
    A second imaging step of acquiring one or more second images by imaging at a resolution higher than that of the first imaging step for each of the regions of interest set in the region of interest setting step;
    A photographing condition setting step of setting a photographing condition based on the one or more second images acquired in the second photographing step;
    And a third imaging step of performing imaging of the same resolution as the first imaging step with respect to the first visual field range based on the imaging condition set in the imaging condition setting step.
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