JP2018146447A - Stirring micro object observation device, and stirring micro object observation method - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a stirring micro object observation device capable of suppressing a reflectance of a stirring blade or a stirring probe at the imaging time and an image degradation caused by reflection light of illumination by the stirring blade or the stirring prove and the reflectance of the scattered light and capable of suppressing a blurred image caused by the movement of an observation object to the minimum possible limit.SOLUTION: A stirring micro object observation device includes: imaging means for imaging a micro object contained in the liquid stirred inside a vessel from the bottom of the vessel toward an inner part direction of the vessel; irradiation means for irradiating an inner part of the vessel; positional information detection means for detecting a position of the stirring blade or a stirring probe used for stirring the liquid and rotationally moving; and means for determining an imaging period based on the positional information of the stirring blade or the stirring probe detected by the positional information detection means.SELECTED DRAWING: None

Description

  The present invention relates to a stirring minute object observation apparatus and a stirring minute object observation method.

  In general, an apparatus that stirs a liquid in a container using a stirrer called a stirrer, a stirring blade, an impeller, a stirrer, and a rotor is used in various fields.

  In addition, an apparatus for monitoring the state of a solution in an agitating vessel by sensors such as temperature and pH, and environmental control equipment such as a temperature control equipment has been developed.

  For example, measuring the concentration of microorganisms in the liquid by irradiating laser light from the side of the container while stirring the liquid in the container with a stir bar and measuring the fluorescence intensity of the microorganisms stained with the fluorescent material. Has been proposed (see, for example, Patent Documents 1 and 2).

  In addition, when cells are cultured in a stirring vessel, it has a plurality of sensors for detecting the state of the culture solution and a plurality of actuators for operating the culture solution, and the actuator is based on information from the sensor group. It has been proposed to optimize the culture conditions by controlling (see, for example, Patent Document 3).

  Furthermore, for a stirring vessel that performs cell culture using microcarriers, a thin chamber for cell observation protruding from the peripheral side of the vessel is provided, and a portion of the culture solution is moved to the thin chamber by tilting the vessel during cell observation It has been proposed to photograph cells in the culture solution with a CCD camera (see, for example, Patent Document 4).

  In recent years, such an apparatus is used for a method of culturing in large quantities with stirring in order to disperse cells in a culture solution with the development of antibody drugs and regenerative medicine.

  The present invention minimizes reflection of a stirring blade or stirrer during imaging, image quality deterioration due to reflection of reflected light and scattered light from the stirrer or stirrer, and image blur due to movement of an observation object. An object of the present invention is to provide an agitating minute object observation device that can be suppressed.

  As means for solving the above-mentioned problem, the stirring micro object observation device of the present invention includes an imaging means for imaging a micro object contained in a liquid stirred in a container from the bottom surface of the container toward the inside of the container. Illuminating means for illuminating the interior of the container; position information detecting means for detecting the position of a rotating stirring blade or stirring bar used for stirring the liquid; and the stirring blade detected by the position information detecting means or And means for determining a timing for imaging based on the position information of the stirrer.

  According to the present invention, reflection of a stirring blade or a stirring bar at the time of imaging, image quality deterioration due to reflection of reflected light and scattered light from the stirring blade or the stirring bar, and image blur due to movement of an observation object are minimized. It is possible to provide an agitation minute object observation device that can be suppressed.

FIG. 1 is a schematic diagram showing an example of a stirring minute object observation apparatus of the present invention. FIG. 2 is a plan view showing an example of the shape of a stirring blade that can be used in the stirring minute object observation apparatus shown in FIG. FIG. 3 is a plan view showing another example of the shape of the stirring blade that can be used in the stirring minute object observation apparatus shown in FIG. FIG. 4 is a plan view illustrating another example of the shape of the stirring blade that can be used in the stirring microscopic object observation apparatus illustrated in FIG. 1. FIG. 5 is a block diagram illustrating an example of the stirring minute object observation apparatus illustrated in FIG. 1. 6 is a cross-sectional view taken along line AA showing an example of the stirring minute object observation apparatus shown in FIG. FIG. 7 is a cross-sectional view taken along line AA showing another example of the stirring minute object observation apparatus shown in FIG. FIG. 8 is a schematic diagram showing another example of the stirring minute object observation apparatus of the present invention. FIG. 9 is a plan view showing an example of the shape of a stirring bar that can be used in the stirring minute object observation apparatus shown in FIG. FIG. 10 is a plan view showing another example of the shape of the stirring bar that can be used in the stirring minute object observation apparatus shown in FIG. FIG. 11 is a plan view showing an example of a rotor that can be used in the stirring minute object observation apparatus shown in FIG. FIG. 12 is a cross-sectional view taken along the line C-C showing an example of a positional relationship between a rotor usable in the stirring minute object observation apparatus shown in FIG. 8 and position information detecting means. FIG. 13 is a cross-sectional view showing an example of illumination means that can be used in the stirring minute object observation apparatus shown in FIG. FIG. 14 is a block diagram illustrating an example of the stirring minute object observation device illustrated in FIG. 8. FIG. 15 is a cross-sectional view taken along line BB showing an example of the stirring minute object observation apparatus shown in FIG. 16 is a cross-sectional view taken along line BB showing another example of the stirring minute object observation apparatus shown in FIG.

(Stirring minute object observation device and stirring minute object observation method)
The stirring minute object observation device of the present invention includes an imaging unit that images a minute object contained in a liquid stirred in a container from the bottom surface of the container toward the inside of the container, and an illumination unit that illuminates the inside of the container. And a position information detecting means for detecting the position of a rotating stirring blade or stirring bar used for stirring the liquid, and a timing for imaging based on the position information of the stirring blade or stirring bar detected by the position information detecting means And a means for determining the other, and other means as necessary.
The stirring minute object observing method of the present invention includes an imaging process for imaging from the bottom surface of the container toward the inside of the container, an illumination process for proving the inside of the container, and a rotating stirring blade used for stirring the liquid. Or a position information detection step for detecting the position of the stirrer, and a step for determining a timing for imaging based on the position information of the stirrer blade or the stirrer detected by the position information detection step. These steps are included.

In the conventional observation method, the stirring minute object observation apparatus and the stirring minute object according to the present invention can be used to observe microorganisms, cells, and minute objects from the side of the container using an optical microscope. This is based on the knowledge that there is a problem that the image quality deteriorates because the object to be photographed is distorted due to the influence of the thickness and the side surface being curved.
In addition, when the diameter of the cylindrical container is large or when the stirring speed is high, the flow velocity near the wall surface is fast, so that the conventional method of observing from the side surface of the container blurs the image even if the exposure time is shortened as much as possible. For this reason, in order to shorten the exposure time and to capture an object to be photographed with appropriate exposure, when introducing strobe shooting with high intensity in the shooting area, it is necessary to use laser light from the viewpoint of shortening the pulse and increasing the intensity. is there. However, the use of the laser beam is based on the knowledge that there is a problem that it is generally difficult to use because the object to be photographed is damaged.
Furthermore, with the conventional observation method, the image of the stirring blade is reflected in the image, the light intensity varies greatly due to the scattering of illumination light by the stirring blade, and whiteout occurs, and the object in the stirring state cannot be observed. It is based on the knowledge that there is.

  The minute object means, for example, a particle having a diameter of 1 μm or more and 1,000 μm or less, and examples include living organisms (microorganisms), animal cells, plant cells, and fine particles.

  The fine particles are not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include inorganic or organic compound particles. Examples of the inorganic or organic compound particles include toner.

<Imaging means and imaging process>
The imaging means is means for imaging a minute object contained in the liquid stirred in the container from the bottom surface of the container toward the inside of the container.
The imaging step is a step of imaging a minute object included in the liquid stirred in the container from the bottom surface of the container toward the inside of the container. The imaging step can be suitably performed by the imaging unit.
Examples of the imaging means include a microscope unit.

  The microscope unit includes an objective lens, a lens barrel, and a camera, and may further include other members as necessary.

The objective lens is not particularly limited and may be appropriately selected depending on the purpose. For example, in a configuration using a lens barrel provided with an imaging lens, an infinite objective lens and an imaging lens are provided. In a configuration using a lens barrel that is not, a finite objective lens or the like can be cited.
There is no restriction | limiting in particular as a kind of said objective lens, According to the objective, it can select suitably, For example, the container in which the stirring solution of the observation target object and the lens which correct | amended the aberration by a stirring solution are preferable. In the case where a lens in which the aberration is corrected is not used, it is preferable to take an image in an area close to the bottom of the container in order to minimize deterioration in image quality due to aberration.

There is no restriction | limiting in particular as said camera, According to the objective, it can select suitably, For example, using a camera provided with CCD (Charge-Coupled Device), CMOS (Complementary Metal Oxide Semiconductor) etc. in an image sensor. it can. Further, when a CMOS is used as the image sensor, it is preferable to use a global shutter type camera.
There is no restriction | limiting in particular as said global shutter system camera, According to the objective, it can select suitably, For example, GO-5000C-USB (made by JAI Corporation) etc. are mentioned.
There is no restriction | limiting in particular as a magnitude | size of the said image pick-up element, According to the objective, it can select suitably.
There is no restriction | limiting in particular as the number of pixels of the said image pick-up element, According to the objective, it can select suitably.
There is no restriction | limiting in particular as said camera, According to the objective, a shape, a magnitude | size, a structure, a material, etc. can be selected suitably.
The size of the camera is not particularly limited as long as it can fit in the housing 10 and can be appropriately selected according to the purpose.
The optical magnification of the image pickup means is not particularly limited, but is preferably set based on the ratio of the area of the image pickup device of the camera to the area to be imaged. In the case of observation, the objective lens and the imaging lens are selected and configured so that the optical magnification is four times.

<Illumination means and illumination process>
The illuminating means is not particularly limited, as long as it does not affect the stirring minute object of the imaging target, can secure the sensitivity of the camera, and can obtain a sufficient contrast in the transmitted light image. Can be appropriately selected according to the above, and examples thereof include a halogen lamp and an LED lamp.
The illumination process is a process of illuminating the inside of the container. The illumination step can be preferably performed by the illumination unit.
There is no restriction | limiting in particular as said illumination means, According to the objective, a shape, a magnitude | size, a structure, a material, etc. can be selected suitably.
There is no restriction | limiting in particular as a shape of the said illumination means, According to the objective, it can select suitably, For example, circular, square, a rectangle etc. are mentioned.

<Position information detection means and position detection step>
The position information detecting means is means for detecting the position of a rotating stirring blade or stirring bar used for stirring.
The position information detecting step is a step of detecting the position of a rotating stirring blade or stirring bar used for stirring. The position information detecting step can be preferably performed by the position information detecting means.
The position information detecting means is not particularly limited as long as it can detect the position information of the stirring blade or the stirring bar that rotates in the stirring vessel, and can be appropriately selected according to the purpose. A method of optically detecting using a sensor, a transmission type photosensor, a method of obtaining information of a rotation angle using a mechanical switch from a mechanism for rotational movement, and a stirring blade or a stirring bar using a camera. Means for capturing a motion and detecting the position of the stirring blade or the stirring bar from the image can be used.

  There is no restriction | limiting in particular as said reflection type photosensor, According to the objective, it can select suitably, For example, reflection type photo interrupter SG-2BC (made by Kodenshi Co., Ltd.) etc. are mentioned.

  There is no restriction | limiting in particular as said transmissive | pervious photosensor, According to the objective, it can select suitably, For example, transmissive | pervious photointerrupter SG206 (made by Kodenshi Co., Ltd.) etc. are mentioned.

<Means for Determining Image Timing Based on Position Information of Stirrer Blade or Stirrer, and Step of Determining Time to Image Based on Position Information of Stirrer Blade or Stirrer>
The means for determining the timing for imaging based on the position information of the stirring blade or the stirring bar is a means for outputting a signal for the imaging means to acquire an image of the position information of the stirring blade or the stirring bar.
The step of determining the timing of imaging based on the position information of the stirring blade or the stirring bar is a step of outputting a signal for the imaging means to acquire an image of the position information of the stirring blade or the stirring bar.
The step of determining the image capturing time based on the position information of the stirring blade or the stirring bar can be suitably performed by means for determining the time of image capturing based on the position information of the stirring blade or the stirring bar.
Examples of the means for determining the timing of imaging based on the position information of the stirring blade or the stirring bar include a pulse generator and a microcontroller.

  Hereinafter, specific embodiments of the stirring micro object observation device of the present invention will be described in detail with reference to the drawings, and the stirring micro object observation method of the present invention will be described.

<First embodiment>
FIG. 1 is a schematic diagram of an example of a stirring micro object observation apparatus according to the present invention, and FIGS. 2 to 4 are plan views showing shapes of stirring blades that can be used in the stirring micro object observation apparatus shown in FIG. FIG. 5 is a block diagram showing an example of the stirring minute object observation device shown in FIG. 1, and FIGS. 6 and 7 show the stirring blade and position information detection when the stirring minute object stirring device shown in FIG. 1 is used. It is sectional drawing in the AA line which shows an example of the positional relationship of a means and an opening part. With reference to these drawings, a stirring minute object observation method using the stirring minute object observation apparatus of FIG. 1 will be described.

The stirring minute object observation apparatus 1 includes a housing 10 and a transparent container 20, and includes a microscope unit 50 that is an imaging unit inside the housing 10, and the transparent container 20 is disposed on an upper bottom surface of the housing 10. The illumination light source 200 that is installed and illuminates the inside of the transparent container 20 is located above the transparent container 20 and directly above the microscope unit 50.

There is no restriction | limiting in particular as the said housing | casing 10, According to the objective, a shape, a structure, a magnitude | size, and a material can be selected suitably.
The shape of the casing 10 is not particularly limited as long as the microscope unit 50 accommodated therein and the transparent container 20 loaded on the upper side of the casing 10 can be stably installed, and are appropriately selected according to the purpose. For example, a cubic shape and a rectangular parallelepiped shape are preferable.
The structure of the casing 10 includes a cavity for accommodating the microscope unit 50 therein, an opening 11 for imaging the inside of the transparent container 20 loaded on the upper portion by the microscope unit 50, and position information of the stirring blade 21 The reflection type photosensor 300 is provided as position information detection means for detecting the above. The reflective photosensor 300 is a sensor including a light emitting element 302 and a light receiving element 303, and is attached to the upper bottom surface of the housing 10 as shown in FIG. Further, by configuring a peripheral circuit as shown in FIG. 5, light emitted from the light emitting element 302 is reflected, light reflected by the light receiving element 303 is received, and the light can be output as a voltage signal 401. .
The size of the casing 10 is not particularly limited as long as the imaging unit and the position information detection unit can be accommodated therein, and can be appropriately selected according to the purpose.
The material of the housing 10 is not particularly limited as long as it can withstand the load applied to the housing 10 and can be appropriately selected according to the purpose.

  The transparent container 20 contains a culture solution 25 containing microorganisms, and includes a stirring blade 21 for stirring the culture solution 25, a motor 24 for rotating the stirring blade 21, and a shaft 23 for transmitting the power. In addition, by stirring the culture solution 25 with the stirring blade 21 that rotates, the microorganisms in the culture solution 25 can be cultured without being precipitated.

There is no restriction | limiting in particular as the said transparent container 20, According to the objective, a shape, a structure, a magnitude | size, and a material can be selected suitably.
The shape of the transparent container 20 is not particularly limited as long as it does not hinder the installation of the shaft 23 for transmitting power to the stirring blade 21 and can be appropriately selected according to the purpose. Can be mentioned.
There is no restriction | limiting in particular as a structure of the said transparent container 20, Although it can select suitably according to the objective, For example, it does not need to be transparent as a whole, and it can be set as the structure which has a partially transparent part.
There is no restriction | limiting in particular as a material of the said transparent container 20, The material which can permeate | transmit the wavelength of the component required for observation among illumination light can be used, Although it can select suitably according to the objective, For example, glass And quartz.

  For example, a cylindrical beaker is preferable as the transparent container 20.

There is no restriction | limiting in particular as the said stirring blade 21, According to the objective, a shape, a structure, a magnitude | size, and a material can be selected suitably.
There is no restriction | limiting in particular as a shape of the said stirring blade 21, Although it can select suitably according to the objective, For example, a linear shape without a branch like FIG. 2, or like FIG.3 and FIG.4 The thing of the shape branched to the trident or the four fork centering on the shaft is mentioned.
The material of the stirring blade 21 preferably has a metallic luster because it has corrosion resistance and is detected by the position information detecting means. Examples of the material include stainless steel.

  The microscope unit 50 includes at least an objective lens 51, a lens barrel 52, and a camera 53, and the objective lens 51 is installed toward the transparent container 20 from the opening 11 vacated immediately above the microscope unit 50.

The objective lens 51 is not particularly limited and can be appropriately selected according to the purpose. For example, in a configuration using a lens barrel provided with an imaging lens, an infinite objective lens or imaging lens is used. In a configuration using a lens barrel that is not provided, a finite objective lens or the like may be used.
As the lens barrel provided with the imaging lens, a commercially available product can be used, and examples of the commercially available product include a small incident light microscope CM series manufactured by Nikon Corporation.
A commercially available product can be used as the infinite objective lens. Examples of the commercially available product include Nikon Corporation, bright field objective lens CF IC EPI Plan series, and the like.
There is no restriction | limiting in particular as said finite system lens, According to the objective, it can select suitably, For example, the objective lens based on DIN specification etc. are mentioned.

Next, a first embodiment of the stirring minute object observation method of the present invention will be described with reference to FIGS. As shown in FIG. 5, the reflective photosensor 300 is connected to a power supply voltage 304 (+ 5V, etc.), a resistor 305, and a resistor 306. The resistor 305 and the resistor 306 respectively adjust the light amount of the light emitting element 302 and receive light. The element 303 represents a resistance necessary for adjusting the light reception signal level. In FIG. 6, the reflective photosensor 300 as position information detection means faces the lower bottom surface of the transparent container 20, and the moment when the rotating stirring blade 21 is positioned above the reflective photosensor 300. In addition, the light emitted from the light emitting element 302 is reflected by the stirring blade 21, and the reflected light is detected by the light receiving element 303. When the light receiving element 303 detects light, a voltage signal 401 is output, and the voltage signal 401 is input to the pulse generator 40. That is, when the stirring blade 21 is positioned above the reflective photosensor 300, the voltage signal 401 is output at a predetermined value or more (more than a threshold value), and the trigger signal A ₁ 402 that determines the timing of imaging from the pulse generator 40 is displayed on the camera 53. The camera 53 performs an imaging operation and acquires image data 403. When the camera 53 acquires the image data 403, the image data 403 is transmitted to the computer 60 using a transmission path such as a USB (Universal Serial Bus) cable or a LAN (Local Area Network) cable and stored.

  The imaging time means a time when the position information detecting means detects that the stirring blade or the stirring bar is located at a specific place. In the first embodiment, the position information detection means detects that the stirring blade or the stirring bar is located at a specific location. The position information detection means blocks the light from the light source by the stirring blade or the stirring bar. The imaging means means a time when the light from the light source is not blocked. The timing for imaging varies depending on the type of the stirring blade and the position where the position information detecting means is attached. For example, when the stirring blade 21 having the shape shown in FIG. 2 is used as the time for imaging, the position of the opening 11 on the upper bottom surface of the housing 10 and the reflection type photosensor as shown in FIGS. The position 300 can be arranged at a position where the opening 11, the center of the bottom surface of the transparent container, and the central angle of the sector formed by the reflective photosensor 300 are 90 °.

  With this configuration, as shown in FIG. 7, when the stirring blade 21 is positioned above the reflective photosensor 300, the reflective photosensor 300 detects the stirring blade 21 and passes the pulse generator 40 to the camera. A signal is output to 53. For this reason, since the stirring blade 21 is positioned above the reflective photosensor 300 at the time of imaging, the stirring blade 21 blocks the illumination light, the reflected image of the stirring blade 21 appears in the captured image, and It is possible to suppress deterioration in image quality due to reflection of reflected light and scattered light from the stirring blade 21. In addition, since the timing for performing the imaging can be determined by the stirring blade 21, the conditions at the time of acquiring the image data 403 can be easily controlled, and stable data can be acquired.

<Second Embodiment>
FIG. 8 is a schematic diagram showing another example of the stirring minute object observation apparatus of the present invention, and FIGS. 9 to 10 show examples of the shape of the stirring bar usable in the stirring minute object observation apparatus shown in FIG. FIG. 11 is a plan view showing an example of a rotor that can be used in the stirring minute object observation device shown in FIG. 8, and FIG. 12 shows the rotor and position information detection means shown in FIG. FIG. 13 is a plan view showing an example of illumination means that can be used in the stirring minute object observation device shown in FIG. 8, and FIG. 13 is a cross-sectional view taken along the line CC indicating the positional relationship with the transmission type photosensor. FIG. 15 is a block diagram illustrating an example of the stirring minute object observation device illustrated in FIG. 8, and FIGS. 15 to 16 illustrate stirring blades, position information detection means, and openings when the stirring minute object observation device illustrated in FIG. 8 is used. In the BB line showing an example of the positional relationship of the parts It is a surface view. The stirring minute object observation method using the stirring minute object observation apparatus in FIG. 8 will be described with reference to these drawings. In the second embodiment, the same reference numerals as those in the first embodiment can be used unless otherwise specified.

  The stirring minute object observation apparatus 1 includes a housing 10 and a transparent container 20, and as shown in FIGS. 8 and 13, as an illumination unit 201 for illuminating the inside of the transparent container on the peripheral side surface of the housing 10. A ring-type LED light source 203 having an LED light source 204 is provided.

  A commercially available product can be used as the ring-type LED light source 203. Examples of the commercially available product include LED lighting DR-LA series (manufactured by Nisshin Electronics Industry Co., Ltd.).

There is no restriction | limiting in particular as the said housing | casing 10, According to the objective, it can select suitably about a shape, a structure, a magnitude | size, and a material.
The structure of the housing 10 is a transmission type as position information detecting means for detecting position information of the rotor 70 connected to the housing 10 by the rotation shaft 73 on the same bottom surface as the opening 11 and the stirrer 22. A photo sensor 301 is included.

The transparent container 20 contains a culture solution 26 containing cells, and a stirrer 22 for stirring the culture solution 26 is placed in the culture solution 26. The transparent container 20 is preferably provided with a lid 27 for the purpose of preventing contamination of the culture solution 26 due to factors outside the container such as falling bacteria.
The shape, structure, size, and material of the lid 27 are not particularly limited and can be appropriately selected according to the purpose. To the captured image by a lighting fixture or the like installed on the top of the stirring minute object observation device 1 For the purpose of preventing the influence of the above, it is preferable to use an opaque material for the lid 27 or attach an opaque seal to the lid 27.
In the present embodiment, the transparent container 20, the stirrer 22, and the lid 27 are separately prepared and combined. However, the transparent container 20, the stirrer, and the lid are integrated. The container may be used.
Commercially available products can be used as containers having a structure in which the transparent container, the stirring bar, and the lid are integrated. Examples of the commercially available products include a single-use bioreactor for culturing iPS cells (ABLE). Etc.).

  As shown in FIG. 11, the rotor 70 has a flat disk shape and a gear shape having a tooth shape on the peripheral side surface thereof. The rotor 70 rotates when the power of a motor 71 provided in the housing 10 is transmitted from a pinion gear 72. As shown in FIG. 8, the rotor 70 is located between the upper bottom surface of the housing 10 and the microscope unit 50.

As shown in FIG. 11, the structure of the rotor 70 has two openings 75, slits 74, and magnets 76 around the rotation shaft 73 in two points symmetrically. The opening 75 is a circular hole that penetrates the rotor 70, and is opened for the microscope unit 50 installed in the housing 10 to image the inside of the transparent container 20.
There is no restriction | limiting in particular as a magnitude | size of the said rotor 70, According to the objective, it can select suitably.
There is no restriction | limiting in particular as a material of the said rotor 70, According to the objective, it can select suitably.

  The slit 74 is opened to detect position information by the transmissive photosensor 301. As shown in FIG. 11, the center of the opening 75 is the center of the opening 11 opened in the housing 10. The overlapping position can be detected, and the mounting positions of the slit 74 and the transmissive photosensor 301 can be adjusted so that the influence on the imaging of the stirrer 22 is minimized.

The stirring element 22 in the transparent container 20 is transmitted with power rotating by the magnet 76 of the rotor 70. Therefore, since the stirring bar 22 always rotates according to the position of the magnet 76 of the rotor 70, the stirring bar 22 can be prevented from overlapping with the slit 74 and the opening 75, so that the stirring bar 22 can be used at the time of imaging. Can be prevented from being positioned on the objective lens 51, and the reflection of the stirrer 22 into the captured image can be prevented.
There is no restriction | limiting in particular as a shape, a structure, a magnitude | size, and a material of the said stirring element 22, Although it can select suitably according to the objective, It is preferable to have magnetism.

  As shown in FIG. 12, the transmissive photosensor 301 has a light emitting element 302 and a light receiving element 303 facing each other, and the rotor 70 is partially accommodated in a U-shaped groove of the transmissive photosensor 301. It is arranged so that. Normally, the light from the light emitting element 302 is not received by the light receiving element 303 by the rotor 70, but when the rotor 70 rotates and the slit 74 is positioned on the optical axis of the light emitting element 302, the light receiving element 303 is moved to the light emitting element 302. Light is detected as a voltage signal 401.

Next, a second embodiment of the stirring microscopic object observation method of the present invention will be described with reference to FIGS. As shown in FIG. 14, the transmissive photosensor 301 is connected to a power supply voltage 304 (+5 V, etc.), a resistor 305, and a resistor 306, and the resistor 305 and the resistor 306 respectively adjust the light amount of the light emitting element 302 and receive light. The element 303 represents a resistance necessary for adjusting the light reception signal level. 15 and 16, the transmissive photosensor 301 serving as position information detection means is a moment when the slit 74 is positioned on the optical axis of the light emitting element 302 and the light receiving element 303 arranged in the groove of the transmissive photosensor 301. In addition, position information is detected. When the light receiving element 303 detects light, a voltage signal 401 is output, and the voltage signal 401 is input to the pulse generator 40. That is, when the slit 74 is positioned on the optical axis of the transmissive photosensor 301, the voltage signal 401 is output at a predetermined value or more (a threshold value or more), and the trigger signal A ₁ 402 for determining the imaging timing from the pulse generator 40 is output. A trigger signal A ₂ 404 is generated for causing the camera 53 to emit light at the moment of imaging, and the trigger signal B 405 is output to the ring-type LED light source 203 by the illumination control device 202 to cause strobe light emission. Here, the timing at which the trigger signal A ₁ 402 and the trigger signal A ₂ 404 are generated is not the same, the time interval from when the camera 53 receives the trigger signal A ₁ 402 until the actual imaging is performed, and the trigger signal A _2. The lighting control device 202 having received 404 can be adjusted in advance in consideration of the difference from the time interval until the ring-type LED light source 203 is turned on.
The camera 53 of the microscope unit 50 receives the trigger signal A ₁ 402 to perform an imaging operation, and transfers the image data 403 through a transmission path such as a USB (Universal Serial Bus) cable or a LAN (Local Area Network) cable. Used to transmit to the computer 60 for storage.

  The imaging time means a time when the position information detecting means detects that the stirring blade or the stirring bar is located at a specific place. The time when the position information detecting means in the second embodiment detects that the stirring blade or the stirring bar is located at a specific location is the time when the slit 74 of the rotor 70 is the position information detecting means of the transmission type photosensor 301. Means the time of the location. The timing for imaging varies depending on the type of the stirring bar and the mounting position of the position information detecting means. For example, when the stirrer 22 having the shape shown in FIG. 9 is used, the position of the opening 11 on the upper bottom surface of the housing 10 and the position of the stirrer 22 as shown in FIGS. The position with respect to the end point may be arranged at a position where the center angle of the sector formed by the opening 11, the center of the bottom surface of the transparent container, and the end point of the stirrer 22 is 90 °.

  By adopting such a configuration, it is possible to suppress the stirrer from blocking the illumination light, the reflection of the stirrer in the captured image, and the deterioration of the image quality due to the reflection of the reflected light and scattered light of the stirrer. can do. In addition, since the illumination means is controlled by the position information of the stirrer, the stirring minute object can be imaged with appropriate exposure even if the exposure time is shortened when the dynamics of the stirring minute object in the solvent is intense, Stable image data can be acquired by suppressing the reflection of the stirrer and image blur.

As an aspect of this invention, it is as follows, for example.
<1> Imaging means for imaging a minute object contained in the liquid stirred in the container from the bottom surface of the container toward the inside of the container;
Illumination means for illuminating the interior of the container;
Position information detection means for detecting the position of a rotating stirring blade or stirring bar used for stirring the liquid;
A stirring microscopic object observing apparatus, comprising: means for determining a timing for imaging based on positional information of the stirring blade or the stirring bar detected by the position information detecting means.
<2> The stirring fine object observation device according to <1>, wherein the imaging unit is a microscope unit.
<3> The stirring microscopic object observation apparatus according to <2>, wherein the microscope unit includes an objective lens, a lens barrel, and a camera.
<4> The stirring minute object observation device according to <3>, wherein the objective lens is at least one of a finite system lens and an objective lens including a finite system lens, an infinite system lens, and an imaging lens.
<5> The stirring microscopic object observation apparatus according to any one of <3> to <4>, wherein the imaging element provided in the camera is at least one of a CCD and a CMOS.
<6> The stirring fine object observation device according to any one of <1> to <5>, wherein the illumination unit irradiates light from a side surface of the container toward a lower bottom surface side.
<7> The stirring fine object observation device according to any one of <1> to <6>, wherein the illumination unit is at least one of a halogen lamp and an LED lamp.
<8> The stirring fine object observation device according to any one of <1> to <7>, wherein the position information detection unit is a photosensor.
<9> The position information detection means detects the position of the stirring blade or the stirring bar from the image by capturing the motion of the stirring blade or the stirring bar using a reflection type photo sensor, a transmission type photo sensor, a mechanical switch, and a camera. The stirring microscopic object observing device according to <1> to <8>, wherein the stirring microscopic object observation device is at least one selected from a method for performing the above.
<10> An imaging step of imaging a minute object included in the liquid stirred in the container from the bottom surface of the container toward the inside of the container;
An illumination process for illuminating the interior of the container;
A position information detecting step for detecting the position of a rotating stirring blade or stirring bar used for stirring the liquid;
And a step of determining a timing for imaging based on the position information of the stirring blade or the stirring bar detected by the position information detecting step.

  According to the stirring minute object observation device according to any one of <1> to <9> and the stirring minute object observation method according to <10>, the problems in the related art are solved, and the object of the invention Can be achieved.

JP 2014-42463 A JP 2014-55796 A Special table 2008-526203 gazette JP 2014-200241 A

DESCRIPTION OF SYMBOLS 1 Stirring minute object observation apparatus 10 Case 21 Stirring blade 22 Stirrer 25 Culture solution containing microorganisms 26 Culture solution containing cells 200 Illumination light source 202 Illumination control device 203 Ring LED light source 300 Reflective photosensor 301 Transmission Type photosensor 302 light emitting element 303 light receiving element 40 pulse generator 50 microscope unit 51 objective lens 52 lens barrel 53 camera

Claims (4)

Imaging means for imaging a minute object contained in the liquid stirred in the container from the bottom surface of the container toward the inside of the container;
Illumination means for illuminating the interior of the container;
Position information detection means for detecting the position of a rotating stirring blade or stirring bar used for stirring the liquid;
A stirring micro-object observation apparatus, comprising: means for determining a timing for imaging based on position information of the stirring blade or the stirring bar detected by the position information detecting means.   The stirring microscopic object observation apparatus according to claim 1, wherein the illumination unit irradiates light from a side surface of the container toward a lower bottom surface side.   The stirring fine object observation apparatus according to claim 1, wherein the position information detection unit is a photosensor. An imaging step of imaging a minute object contained in the liquid stirred in the container from the bottom surface of the container toward the inside of the container;
An illumination process for illuminating the interior of the container;
A position information detecting step for detecting the position of a rotating stirring blade or stirring bar used for stirring the liquid;
A method for observing a stirring minute object, comprising: a step of determining an image capturing time based on position information of the stirring blade or the stirring bar detected by the position information detecting step.