WO2018158957A1 - Système d'observation de cellules - Google Patents

Système d'observation de cellules Download PDF

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Publication number
WO2018158957A1
WO2018158957A1 PCT/JP2017/008619 JP2017008619W WO2018158957A1 WO 2018158957 A1 WO2018158957 A1 WO 2018158957A1 JP 2017008619 W JP2017008619 W JP 2017008619W WO 2018158957 A1 WO2018158957 A1 WO 2018158957A1
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WO
WIPO (PCT)
Prior art keywords
image
cell observation
data
observation system
unit
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PCT/JP2017/008619
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English (en)
Japanese (ja)
Inventor
倫誉 山川
快彦 岩尾
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株式会社島津製作所
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Priority to PCT/JP2017/008619 priority Critical patent/WO2018158957A1/fr
Publication of WO2018158957A1 publication Critical patent/WO2018158957A1/fr

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    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
    • G06T1/00—General purpose image data processing
    • H—ELECTRICITY
    • H04—ELECTRIC COMMUNICATION TECHNIQUE
    • H04N—PICTORIAL COMMUNICATION, e.g. TELEVISION
    • H04N1/00—Scanning, transmission or reproduction of documents or the like, e.g. facsimile transmission; Details thereof

Definitions

  • the present invention relates to a cell observation system that observes cells using a holographic microscope.
  • a cell observation apparatus (holography observation apparatus) using a holographic microscope.
  • holographic observation apparatus an image of a cell accommodated in a culture plate is acquired while the cell is alive, and various measurements and analyzes are performed based on the image data.
  • a light beam with a uniform phase is divided into two parts, one of which is irradiated to the object to be passed or reflected, and the other is left as it is.
  • an image holographic image or hologram
  • the above-mentioned holographic image is measured by an image sensor for each region obtained by dividing the culture plate into a plurality, and a phase image and an intensity image are created by numerical calculation. Then, the phase images and intensity images of the plurality of divided regions are connected (also referred to as “tiling”) to obtain the phase image and intensity image of the entire culture plate.
  • the system of Patent Document 1 has a configuration in which a cell observation device and a display device are connected to a management server via a communication network.
  • the cell observation apparatus includes an optical microscope, an image sensor (CCD camera) that acquires an image of an object (specimen slide) located in an observation field of the optical microscope, and a tiling image acquired by the image sensor.
  • An image generation unit that generates an image of the entire object is provided.
  • the image data is sent to a management server via a communication network and stored in a database provided in the management server.
  • the management server creates an image (display image) to be displayed on the display device based on the image data of the object, and stores this data in the database. Then, in response to a transmission request from the display device, display image data is read from the database and transmitted to the display device.
  • the holography observation apparatus can observe the cells accommodated in the culture plate as they are alive, if the processing time related to the creation of the image in the cell observation apparatus is long, the culture plate is in a constant temperature culture chamber ( It takes a long time to leave outside the so-called “incubator”), which is not preferable.
  • the problem to be solved by the present invention is a cell observation system including a cell observation device and a management server connected by a communication network, and reduces the burden of processing executed in the cell observation device and creates an image of an observation target object Is to reduce the processing time required.
  • the present invention made to solve the above problems
  • the cell observation device is a) a light source that emits a coherent luminous flux; b) an irradiation optical system that irradiates the observation target object with the light beam emitted from the light source and interferes with the light beam transmitted or reflected at different positions of the observation target object; c) an image sensor for acquiring an interference image of a light beam transmitted or reflected by the observation object; d) a transmission unit that transmits data of an interference image acquired by the image sensor to the management server via the communication network, and
  • the management server is e) an image generation unit that generates an image based on the data transmitted from the transmission unit; f) a data storage unit for storing image information generated by the image generation unit; It is characterized by providing.
  • the irradiation optical system irradiates the light beam on the object to be observed and transmits or transmits the light at a different position of the object to be observed.
  • An interference image of the reflected light beam is formed.
  • the transmission unit transmits the interference image data as it is (in a so-called “raw data” state) to the management server through the communication network.
  • the image generation unit generates an image based on the interference image data transmitted from the transmission unit of the cell observation device, and stores the image information in the data storage unit.
  • the image generation unit of the management server may generate a phase image and a light intensity image of the observation target object based on the data transmitted from the transmission unit.
  • the data of the interference image transmitted from the transmission unit of the cell observation device to the management server is so-called hologram data. Therefore, in this configuration, the image generation unit of the management server configures the phase image and the light intensity image of the observation target object from the hologram data.
  • the cell observation system has a configuration in which one cell observation device is connected to one management server, and a plurality of cell observation devices are connected to one management server. be able to.
  • one or a plurality of display devices may be connected to the management server through a communication network.
  • the number of cell observation devices and the number of display devices may or may not be the same.
  • One of the cell observation devices and display devices connected to one management server may be one and the other may be a plurality.
  • the cell observation system includes a display device connected to the management server via the communication network
  • the management server In response to a request from the display device, the management server reads image information from the data storage unit, generates a display image from the image information, and transmits the display image data to the display device through the communication network. It is preferable to provide a display image generation unit. In this case, it is preferable that the display image generation unit reads out only the image information satisfying the request condition from the display device from the data storage unit.
  • the display image generation unit may include a processed image generation unit that generates the display image by processing the image generated by the image generation unit based on the image information read from the data storage unit. good.
  • processing refers to processing such as tiling, trimming, and resizing.
  • the display image generation unit generates a partial image that is an image of the whole or a part of the observation target object based on the image information read from the data storage unit.
  • a partial image generation unit may be provided.
  • the display image generation unit includes a magnification change image generation unit that generates an image in which an entire or a partial region of the observation target object is enlarged or reduced based on the image information read from the data storage unit. May be. According to this configuration, an image obtained by enlarging or reducing an image of an area to be measured or analyzed in the observation target object at an appropriate magnification can be displayed on the display device.
  • the image sensor when the image sensor cannot acquire an interference image of the entire observation object at once, the image sensor acquires an interference image for each region obtained by dividing the observation object into a plurality of parts, and manages the data. Will be sent to the server.
  • the data may be collectively transmitted to the management server. It is preferable that the transmission unit transmits the data to the management server via the communication network every time it is acquired.
  • the cell observation device includes a plurality of the image sensors, It is preferable that the transmission unit simultaneously transmits data acquired by each of the plurality of image sensors to the management server via the communication network.
  • a solid-state image sensor such as a CMOS image sensor or a CCD image sensor using a photodiode as a photoelectric conversion element can be used.
  • a solid-state image sensor such as a CMOS image sensor or a CCD image sensor using a photodiode as a photoelectric conversion element
  • the light source is configured to emit light beams having a plurality of different wavelengths, and includes a wavelength changing unit that changes the wavelength of the light beams emitted from the light source. According to this configuration, it is possible to emit a light beam having an appropriate wavelength according to the optical characteristics (absorption wavelength, transmission wavelength, etc.) of the observation target object and irradiate the observation target object.
  • the image sensor of the cell observation device acquires the interference image
  • the data is sent to the management server, and an image is generated there.
  • the burden can be reduced and the processing time for image generation can be shortened.
  • the whole block diagram of the cell observation system which is one Example of this invention.
  • FIG. 1 is a schematic diagram showing the overall configuration of the cell observation system according to the present example.
  • the communication network 1 such as the Internet includes a plurality of cell observation terminals 2A, 2B, 2C (hereinafter referred to as “2” unless the individual cell observation terminals need to be specified). Connected).
  • Each cell observation terminal 2 includes a microscope observation unit 21 and a personal computer (PC) 22 connected to the communication network 1.
  • the cell observation terminal 2 corresponds to the cell observation device of the present invention.
  • a management server 3 (hereinafter abbreviated as “server 3”) and a display device 4 are further connected to the communication network 1.
  • the server 3 is generally a high-performance computer, and includes a receiving unit 31 that receives data transmitted from the cell observation terminal 2, a data storage unit 32 that can store a large amount of data, and the computer.
  • a data processing unit 33 that is embodied by executing installed software is provided as a functional block.
  • the display device 4 includes a browsing computer 41, an input unit 42 such as a mouse and a keyboard attached to the browsing computer 41, and a display unit 43.
  • the browsing computer 41 is a general personal computer, and a standard OS as basic software and various software (for example, communication for performing bidirectional communication with the server 3) operating on the OS.
  • a control program, an input control program for controlling the operation of the input unit 42, and a display control program for controlling the operation of the display unit 43) are installed.
  • the browsing computer 41 includes a storage unit (not shown) that stores data such as a display image transmitted from the server 3.
  • the server 3 and the browsing computer 41 are connected to each other via an intranet (or the Internet) such as a LAN.
  • the connection between the server 3 and the browsing computer 41 is not limited to the intranet, and both may be connected via the communication network 1.
  • the microscopic observation unit 21 includes a holographic microscope made of IHM (In-line Holographic Microscopy). That is, as shown in FIG. 2, the microscopic observation unit 21 includes a light source unit 211, an image sensor 212, and a moving mechanism 214.
  • the light source unit 211 emits a laser diode 211a that emits a plurality of laser beams of different wavelengths, and irradiation that irradiates the culture plate 213 as an observation target object as a coherent light beam having a spread of a minute angle (about 10 degrees).
  • An optical system 211b is provided.
  • the image sensor 212 for example, a CMOS (Complementary Metal Oxide Semiconductor) image sensor is used.
  • the microscopic observation unit 21 may include one image sensor 212, but may include a plurality of image sensors 212. In the present embodiment, a total of four image sensors are provided, two on the front side and the back side in FIG. 2 (only two image sensors on the front side of the paper are shown in FIG. 2).
  • a coherent light beam emitted from the light source unit 211 is applied to a predetermined region of the culture plate 213. Then, the light (object light) in the coherent light beam transmitted through the cell S and the culture plate 213 reaches the image sensor 212 while interfering with the light (reference light) transmitted through the adjacent position of the cell S on the culture plate 213. .
  • an interference image (hologram) of the light transmitted through the cell S and the culture plate 213 and the light transmitted through the adjacent position of the cell S on the culture plate 213 is formed. Is done.
  • the light source unit 211 and the image sensor 212 are integrally moved in the XY direction (for example, the direction perpendicular to and parallel to the paper surface of FIG. 2) by the moving mechanism 214, for example. Thereby, the irradiation area (observation area
  • the PC 22 included in the cell observation terminal 2 is a general personal computer, which is equipped with a standard OS as basic software, and further installed with various software operating on the OS.
  • the PC 22 includes a storage unit 220, and controls the operation of the light source unit 211, such as changing the wavelength of the laser beam emitted from the laser diode 211a, as a functional block realized by executing installed software.
  • the data transmission unit 223 transmits the hologram data of the observation area acquired by the image sensor 212 (two-dimensional light intensity distribution data of the hologram formed on the detection surface of the image sensor 212) together with the identification information of the culture plate 213 to the server. 3 to send.
  • An input unit 23 and a display unit 24 are connected to the PC 22.
  • the receiving unit 31 included in the server 3 receives hologram data with identification information of the culture plate 213 sent from the cell observation terminal 2 for each cell observation terminal, and stores the data.
  • the data processing unit 33 includes an arithmetic processing unit 331 as a functional block, an image tiling unit 332, an image dividing unit 333, an instruction receiving unit 334, and an information reading unit 335.
  • the calculation processing unit 331 reads the hologram data from the data storage unit 32, executes predetermined calculation processing, and acquires phase information, intensity information, and pseudo phase information of each observation region from the hologram data. Based on these pieces of information, three types of observation images (phase image, intensity image, and pseudo phase image) are created. In addition, the arithmetic processing unit 331 executes general processing of image data (for example, image resolution change processing, image enlargement / reduction processing).
  • the image tiling unit 332 trims the observation image obtained by the arithmetic processing unit 331 based on the hologram data of the plurality of observation regions. Then, the trimmed observation images are pasted together (tiling) to create an observation image (tiling image) of the entire culture plate 213. Further, the image dividing unit 333 creates an observation image obtained for a part of the observation region of the culture plate 213 or a partial observation image obtained by dividing the observation image of the entire culture plate 213 into a plurality of pieces. The data of the tiling image created by the image tiling unit 332 and the partial observation image created by the image dividing unit 333 are stored in the data storage unit 32 together with the identification information of the culture plate 213.
  • the instruction receiving unit 334 receives a display image transmission request instruction coming from the browsing computer 41.
  • the information reading unit 335 accesses the data storage unit 32 and reads data and information that meet the conditions according to the instruction received by the instruction receiving unit 334.
  • FIG. 4 shows the flow of data and instructions in the operation when creating image data based on the hologram data obtained in the cell observation terminal 2 or creating display image data in response to an instruction from the browsing computer 41. It is a schematic diagram which shows.
  • the operator operates the input unit 23 to input identification information (measurement date, type of cultured cell, etc.) regarding the culture plate 213 and instruct to start the operation, a predetermined light emitted from the light source unit 211 is given.
  • a laser beam (coherent beam) having a wavelength and a coherent distance is irradiated onto the culture plate 213 set in the microscopic observation unit 21.
  • an interference image (hologram) of the light in the coherent light beam that has passed through the cells S and the culture plate 213 and the light that has passed through the adjacent position of the cells S on the culture plate 213 is formed on the detection surface of the image sensor 212.
  • the hologram two-dimensional light intensity distribution data (hologram data) measured by the image sensor 212 is stored in a data file with identification information of the culture plate 213 created in the storage unit 220 of the PC 22. Is done. Subsequently, the light source unit 211 and the image sensor 212 are moved by the moving mechanism 214, and hologram data of the next observation area is acquired by the image sensor 212 and stored in the data file of the storage unit 220.
  • the data transmission unit 223 reads the data file from the storage unit 220, and the identification information of the cell observation terminal 2 is obtained. Is sent to the server 3.
  • the PC 22 of the cell observation terminal 2 can transmit all data files stored in the storage unit 220 to the server 3, but only the data file of the culture plate 213 corresponding to specific identification information. It is of course possible to transmit to the server 3.
  • the operator can instruct what data file is transmitted from the cell observation terminal 2 to the server 3 by operating the input unit 23. Further, by operating the input unit 42 of the display device 4, the PC 22 of the cell observation terminal 2 may be instructed through the server 3 and the communication network 1.
  • the arithmetic processing unit 331 When the receiving unit 31 of the server 3 receives the data file from the cell observation terminal 2, in the data processing unit 33, the arithmetic processing unit 331 performs phase recovery processing, trimming processing, and tiling processing of the hologram data included in the data file. Execute. As a result, an image of each observation region (original image), an image of the entire culture plate 213, and other images obtained by dividing the culture plate 213 into an appropriate number (for example, 1/4 divided image and 1/2 divided image) are created. The The created image data is stored in the data storage unit 32 together with the identification information of the cell observation terminal 2 and the culture plate 213.
  • an operator who measures and analyzes the cultured cells accommodated in the culture plate 213 designates the types of the cell observation terminal 2 and the culture plate 213 from the input unit 42 of the display device 4, and then observes them.
  • the information reading unit 335 of the server 3 selects the entire observation image or partial observation image of the designated cell observation terminal 2 and culture plate 213 from the image data stored in the data storage unit 32.
  • the browsing computer 41 displays the transmitted image on the display unit 43.
  • the operator operates the input unit 42 of the browsing computer 41 to select a partial region of the image of the culture plate 213 displayed on the display unit 43 and information on the image (resolution, Specify the magnification, image type (phase image, intensity image, pseudo phase image, etc.) and request the creation of a display image.
  • the instruction receiving unit 334 determines information and image data to be read in response to a display image creation request from the browsing computer 41 and outputs the information and image data to the information reading unit 335.
  • the information reading unit 335 accesses the data storage unit 32 to read out necessary information and image data and sends them to the arithmetic processing unit 331.
  • the arithmetic processing unit 331 executes a predetermined process (trimming / enlargement / reduction process) based on the sent information and image data to create a display image.
  • the server 3 transmits the data of the display image to the browsing computer 41.
  • the browsing computer 41 that has received the display image displays the display image on the display unit 43.
  • FIG. 5 schematically shows a display image created by the arithmetic processing unit 331 of the server 3 based on the data acquired by the cell observation terminal 2.
  • the upper left diagram in FIG. 5 shows the relationship between the culture plate 213 and the observation areas of the four image sensors 212.
  • each of the four image sensors 212 acquires interference images of light beams that have passed through the quarter regions a1 to a4 of the entire culture plate 213. Therefore, when the culture plate 213 has six wells, observation images of 1.5 wells are created from the data acquired by one image sensor 212.
  • the image sensor 212 has 160 (12 ⁇ 14) pixels, an image obtained by dividing 1.5 wells into 160 sections is acquired by the image sensor 212 (see the lower left diagram in FIG. 5).
  • An observation image of 1.5 wells is created from the image data of 160 sections obtained in this way, and the observation image of one well is trimmed from the observation image of 1.5 wells. An image can be obtained.
  • An observation image of the entire culture plate 213 is created from the image data acquired by the four image sensors 212.
  • the center and the right part of FIG. 5 show examples of display images created by the arithmetic processing unit 331.
  • the center is an image (divided image) for one section, and the right part is an enlargement of a part of this divided image. An image is shown.
  • the server 3 since the data acquired by the image sensor 212 is transmitted to the server 3 as it is, the time required for data transmission can be shortened. Further, the server 3 processes the data acquired by the image sensor 212 to create an image, thereby reducing the burden of processing executed in the cell observation terminal 2 as compared to creating an image in the cell observation terminal 2. It is possible to shorten the processing time required for image creation.
  • the present invention is not limited to the above-described embodiments, and appropriate modifications and changes can be made.
  • the hologram data of the entire culture plate 213 is collectively transmitted to the server 3.
  • the hologram data may be transmitted to the server 3 as soon as the hologram data is acquired for each observation region.
  • the control unit light source control unit, moving mechanism control unit
  • storage unit of the microscopic observation unit 21 are realized by a personal computer 22 different from the microscopic observation unit 21.
  • a storage unit or a control unit may be incorporated.
  • the image dividing unit 333 divides the observation image of the culture plate 213 created by the image tiling unit 332 to create a partial observation image.
  • the image tiling unit 332 creates the image. May be. That is, the image tiling unit 332 can also create a partial observation image by pasting together hologram data of a plurality of observation regions that constitute a partial region of the culture plate 213.
  • the image dividing unit 333 can be omitted.
  • the cell observation unit 21 includes one image sensor that uses a partial region of the observation target object as an observation field, and moves the image sensor relative to the observation target object, so that the entire observation target object is displayed. You may make it acquire the image of.

Abstract

Un système d'observation de cellule selon la présente invention comprend un dispositif d'observation de cellule 2 et un serveur de gestion 3 qui est connecté au dispositif d'observation de cellule 2 via un réseau de communication 1. Le système est caractérisé en ce que : le dispositif d'observation de cellule 2 comprend une unité de source de lumière 211 qui émet des faisceaux cohérents de lumière, un système optique d'irradiation qui irradie un objet 213 devant être observé avec les faisceaux de lumière émis par l'unité de source de lumière 211 et provoque une interférence avec chacun des autres faisceaux de lumière transmis à travers ou réfléchis aux différentes positions de l'objet, un capteur d'image 212 qui acquiert une image d'interférence des faisceaux de lumière transmis à travers ou réfléchis sur l'objet 213, et une unité de transmission qui transmet des données de l'image d'interférence acquise par le capteur d'image 212 au capteur de gestion 3 via le réseau de communication ; et le serveur de gestion 3 comprend une unité de génération d'image qui génère une image sur la base des données transmises par l'unité de transmission, et une unité de stockage de données qui stocke des informations relatives à l'image générée par l'unité de génération d'image.
PCT/JP2017/008619 2017-03-03 2017-03-03 Système d'observation de cellules WO2018158957A1 (fr)

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2005117640A (ja) * 2003-09-16 2005-04-28 Sysmex Corp 標本画像の表示方法およびそれを用いた検索方法、サーベイランス方法、標本画像の表示システム、標本画像表示用プログラムおよびそのプログラムを記録した記録媒体
JP2015505983A (ja) * 2011-12-02 2015-02-26 シー・エス・アイ・アールＣｓｉｒ 物質解析システム、方法、および装置

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2005117640A (ja) * 2003-09-16 2005-04-28 Sysmex Corp 標本画像の表示方法およびそれを用いた検索方法、サーベイランス方法、標本画像の表示システム、標本画像表示用プログラムおよびそのプログラムを記録した記録媒体
JP2015505983A (ja) * 2011-12-02 2015-02-26 シー・エス・アイ・アールＣｓｉｒ 物質解析システム、方法、および装置

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