WO2016051913A1 - 画像処理方法、制御プログラム、記録媒体および画像処理装置 - Google Patents

画像処理方法、制御プログラム、記録媒体および画像処理装置 Download PDF

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WO2016051913A1
WO2016051913A1 PCT/JP2015/069894 JP2015069894W WO2016051913A1 WO 2016051913 A1 WO2016051913 A1 WO 2016051913A1 JP 2015069894 W JP2015069894 W JP 2015069894W WO 2016051913 A1 WO2016051913 A1 WO 2016051913A1
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Prior art keywords
spheroid
image
image processing
circle
region
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PCT/JP2015/069894
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English (en)
French (fr)
Japanese (ja)
Inventor
治郎 津村
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株式会社Ｓｃｒｅｅｎホールディングス
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Publication of WO2016051913A1 publication Critical patent/WO2016051913A1/ja

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
    • G06T1/00—General purpose image data processing
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
    • G06T5/00—Image enhancement or restoration
    • G06T5/20—Image enhancement or restoration using local operators
    • G06T5/30—Erosion or dilatation, e.g. thinning

Definitions

  • the present invention relates to image processing performed on an image obtained by imaging a spheroid, and more particularly to a technique for distinguishing a spheroid included in an image from another object.
  • Cross-reference to related applications The disclosures in the specification, drawings, and claims of the following Japanese application are incorporated herein by reference in their entirety: Japanese Patent Application No. 2014-199859 (filed on September 30, 2014).
  • a cell to be observed is captured by a CCD camera or the like to be converted into image data, and various image processing techniques are applied to the image data for observation or analysis. It has become like this. That is, the cells are imaged by a camera or the like and used for observation in a state where the cells to be observed are held in an appropriate container called a microwell plate or dish.
  • an image other than the observation object may be reflected in the image.
  • an image of streaky foreign matter caused by fine dust or dirt mixed in the atmosphere, scratches or dirt on the container may appear in the image.
  • Such an image of a foreign substance hinders observation.
  • Patent Document 1 As a technique considering this problem, there is one described in Patent Document 1, for example.
  • an image of a container (well) in a state in which cells are not carried is taken in advance, and a difference from the image in the state in which cells are present is obtained, thereby eliminating images of foreign matters such as scratches and dust. is doing.
  • This invention is made in view of the said subject, and provides the technique which can perform the distinction with a spheroid and a foreign material efficiently and with the outstanding reproducibility by performing image processing to the image which imaged the spheroid.
  • the purpose is to do.
  • the image processing method includes a step of detecting a spheroid-like object including the spheroid among objects in the original image from an original image obtained by capturing the spheroid, and the spheroid-like object.
  • a step of executing a duration process using the structural element is a step of detecting a spheroid-like object including the spheroid among objects in the original image from an original image obtained by capturing the spheroid, and the spheroid-like object.
  • exclusion processing is one of image morphological processing, and means processing for shrinking an object region in an image.
  • dilation processing is one of morphological processing and means processing for expanding an object region in an image.
  • the spheroid forms a substantially circular image in the original image, it can be distinguished from the image of the streak-like foreign material based on its shape characteristics. If the spheroid image and the foreign object image are separated from each other in the original image, the distinction between them is relatively easy. In reality, however, there are many cases in which the two overlap each other.
  • the spheroid region and the foreign material region are distinguished by morphological image processing including erosion processing and delation processing.
  • the size setting of the structural element greatly affects the processing result.
  • the size of the structural element is set by subjective judgment and trial of the operator. However, skill is required to set the optimum size, and the processing results may vary.
  • a circle having a diameter corresponding to the size of the detected spheroid-like object is used as the structural element. According to the new knowledge obtained by the inventor of the present application, the size of the structural element can be easily and automatically set in this way, and the spheroid and foreign matter can be efficiently distinguished by morphological processing. It becomes possible.
  • an image processing apparatus includes an image acquisition unit that acquires an original image obtained by capturing a spheroid, and an image processing unit that performs image processing on the original image.
  • the processing means executes the image processing described above.
  • image processing executeds the image processing described above.
  • the structural element corresponding to the size of the spheroid-like object detected in the original image is applied to the morphological process, so that the spheroid and the foreign substance can be distinguished efficiently.
  • FIG. 1 is a side view showing a schematic configuration of an image processing apparatus according to the present invention. It is a figure which shows the principle of the area
  • FIG. 1 is a side view showing a schematic configuration of an image processing apparatus according to the present invention.
  • This image processing apparatus performs image processing on an imaged unit 1 that images a sample in a liquid injected into a recess called a well W formed on an upper surface of a well plate WP, and the captured image. And an image processing unit 2.
  • the well plate WP is generally used in the fields of drug discovery and biological science.
  • the well plate WP is formed in a cylindrical shape with a substantially circular cross section on the top surface of a flat plate, and the bottom surface is transparent and flat.
  • a plurality of round-bottomed wells W are provided.
  • the number of wells W in one well plate WP is arbitrary, for example, 96 (12 ⁇ 8 matrix arrangement) are used.
  • the diameter and depth of each well W is typically about several mm. Note that the size of the well plate and the number of wells targeted by the imaging unit 1 are not limited to these and may be arbitrary, for example, 384 holes.
  • a predetermined amount of liquid as a medium is injected into each well W of the well plate WP, and cells, bacteria, and the like cultured in the liquid under predetermined culture conditions are set as imaging targets of the imaging unit 1.
  • the medium may be one to which an appropriate reagent has been added, or it may be in a liquid state and gelled after being placed in the well W.
  • the imaging unit 1 includes a holder 11, an illumination unit 12, an imaging unit 13, and a control unit 14.
  • the holder 11 is brought into contact with the peripheral edge of the lower surface of the well plate WP carrying the sample together with the liquid in each well W, and holds the well plate WP in a substantially horizontal posture.
  • the illumination unit 12 is disposed above the holder 11, while the imaging unit 13 is disposed below the holder 11.
  • the control unit 14 has a function of controlling operations of these units.
  • the illumination unit 12 emits illumination light Li toward the well plate WP held by the holder 11.
  • white light is used as the illumination light Li.
  • the illumination unit 12 illuminates the sample in the well W provided on the well plate WP from above.
  • An imaging unit 13 is provided below the well plate WP held by the holder 11.
  • an objective lens 131 is disposed immediately below the well plate WP.
  • the optical axis OA of the objective lens 131 is oriented in the vertical direction, and an aperture stop 132, an imaging lens 133, and an imaging device 134 are further provided in order from top to bottom along the optical axis OA of the objective lens 131.
  • the objective lens 131, the aperture stop 132, and the imaging lens 133 are arranged so that their centers are aligned in a line along the vertical direction, and these constitute the imaging optical system 130 as a unit.
  • the respective parts constituting the imaging unit 13 are arranged in a line in the vertical direction, but the optical path may be folded by a reflecting mirror or the like.
  • the imaging unit 13 can be moved by a mechanical drive unit 141 provided in the control unit 14. Specifically, the mechanical drive unit 141 moves the objective lens 131, the aperture stop 132, the imaging lens 133, and the imaging device 134 that form the imaging unit 13 integrally in the horizontal direction, so that the imaging unit 13 becomes the well. Move horizontally with respect to W. When the imaging target in one well W is imaged, the mechanical drive unit 141 positions the imaging unit 13 in the horizontal direction so that the optical axis of the imaging optical system 130 coincides with the center of the well W.
  • the mechanical drive unit 141 performs focusing of the imaging unit with respect to the imaging target by moving the imaging unit 13 in the vertical direction.
  • the mechanical drive unit 141 includes the objective lens 131, the aperture stop 132, the imaging lens 133, and the imaging device so that the objective lens 131 is focused on the inner bottom surface of the well W where the sample that is the imaging target is present. 134 is moved up and down integrally.
  • the mechanical drive unit 141 moves the illumination unit 12 in the horizontal direction integrally with the imaging unit 13. That is, the illuminating unit 12 is arranged so that the optical center thereof substantially coincides with the optical axis OA of the imaging optical system 130, and interlocks with the imaging unit 13 including the objective lens 131 when it moves in the horizontal direction. To move horizontally.
  • the illumination condition is constant regardless of which well W is imaged, and the imaging condition is favorably maintained.
  • the sample in the well W is imaged by the imaging unit 13. Specifically, light emitted from the illumination unit 12 and incident on the liquid from above the well W illuminates the imaging target. Then, light transmitted downward from the bottom surface of the well W is collected by the objective lens 131, and an image of the imaging target is finally formed on the light receiving surface of the imaging device 134 via the aperture stop 132 and the imaging lens 133. This is received by the light receiving element 1341 of the imaging device 134.
  • the light receiving element 1341 is a one-dimensional image sensor, and converts a one-dimensional image of the imaging target imaged on the surface thereof into an electrical signal. As the light receiving element 1341, for example, a CCD sensor can be used.
  • the image signal output from the light receiving element 1341 is sent to the control unit 14. That is, the image signal is input to an AD converter (A / D) 142 provided in the control unit 14 and converted into digital image data.
  • the digital image data obtained in this way is output to the outside via an interface (I / F) 143.
  • the image processing unit 2 includes a control unit 20 having a CPU 201 that controls the operation of each unit of the apparatus.
  • the control unit 20 also stores and saves a graphic processor (GP) 202 responsible for image processing, an image memory 203 for storing and saving image data, programs to be executed by the CPU 201 and GP 202, and data generated by these.
  • a memory 204 a graphic processor (GP) 202 responsible for image processing
  • an image memory 203 for storing and saving image data
  • programs to be executed by the CPU 201 and GP 202 programs to be executed by the CPU 201 and GP 202, and data generated by these.
  • a memory 204 a memory 204.
  • the CPU 201 may also function as the graphic processor 202.
  • the image memory 203 and the memory 204 may be integrated.
  • control unit 20 is provided with an interface (I / F) 205.
  • the interface 205 is responsible for exchanging information with the user and an external device. Specifically, the interface 205 is connected to the interface 143 of the imaging unit 1 via a communication line, and the CPU 201 transmits a control command for controlling the imaging unit 1 to the imaging unit 1. In addition, image data output from the AD converter 142 of the imaging unit 1 is received.
  • the interface 205 is connected to an input device 21 such as an operation button, an input device such as a mouse, a keyboard, or a tablet, or a combination thereof.
  • An operation input from the user received by the input unit 21 is transmitted to the CPU 201 via the interface 205.
  • a display unit 22 having a display device such as a liquid crystal display is connected to the interface 205.
  • the display unit 22 displays an image corresponding to an image signal given from the CPU 201 via the interface 205 and presents information such as a processing result to the user.
  • the image processing unit 2 having the above-described configuration is substantially the same as the configuration of a general personal computer. That is, a general-purpose computer device can be used as the image processing unit 2 of the image processing device.
  • this image processing apparatus it is possible to image cells, bacteria, cell clumps (spheroids), etc. cultured in each well W of the well plate WP, and the captured images can be used for various observations and analysis. Can be provided.
  • this image processing apparatus is applied to the purpose of imaging spheroids cultured in the culture solution injected into the well W.
  • the spheroids in the culture medium are generally spherical, and the spheroids form a substantially circular image in the captured two-dimensional image.
  • each has a substantially circular image, but the shape and size are not necessarily the same.
  • the captured image includes streaks caused by foreign matters such as fibrous dust and dirt mixed in the culture medium or attached to the surface, and scratches and dirt on the bottom surface of the well W. May be reflected.
  • FIG. 2A to 2C are diagrams showing the principle of the region separation processing in the present embodiment.
  • This area separation process is a process for separating an object in which an image of a spheroid and a foreign object are integrated into an spheroid area and a foreign object area in the image.
  • FIG. 2A is a schematic diagram illustrating an example of an image in which a spheroid and a foreign object overlap.
  • An image object OB captured by overlapping a spheroid and a foreign object has a spheroid area Ra having a shape close to a circle corresponding to the spheroid and a foreign object area Rb corresponding to a streak-like foreign object connected to the spheroid.
  • region Rb may contact
  • the difference between the spheroid region Ra and the foreign matter region Rb is obvious, but in the actual image, the boundary between the spheroid region Ra and the foreign matter region Rb in the object OB is determined from the image content. It ’s hard.
  • the morphological processing of the image is known, a detailed description is omitted.
  • the diameter De of the structural element is too large, the shape reproducibility of the spheroid region Ra is deteriorated, and the small spheroid region Ra may be erased in the erosion process.
  • the size of the structural element is determined by subjective judgment of an expert, or the size is changed in multiple stages. To find the optimal size.
  • Such work requires a great deal of labor, and variations in processing results are unavoidable.
  • a manual response is not realistic.
  • the present inventor conducted experiments using various cells forming spheroids and obtained the following knowledge. That is, with respect to an image object having a spheroid region Ra and a foreign object region Rb associated therewith, a morphological process using a structural element having a size calculated based on a predetermined rule from the size of the spheroid region Ra is applied.
  • the region Ra and the foreign material region Rb can be distinguished well.
  • the spheroid region Ra is regarded as a circle
  • an erosion process and a delation are performed using a circle having a diameter not less than (1/4) and not more than (1/3) of the diameter of the circle as a structural element Processing is performed in order.
  • the shape reproducibility of the spheroid region Ra is good, and it is possible to effectively erase the spheroid region Ra while suppressing the residual foreign material region Rb.
  • the image processing of the present embodiment for distinguishing the spheroid region and the foreign material region based on such knowledge will be specifically described with reference to FIGS. 3, 4, 5 ⁇ / b> A to 5 ⁇ / b> D.
  • FIG. 3 is a flowchart showing image processing in this embodiment.
  • FIG. 4 is a flowchart showing the region separation process.
  • FIGS. 5A to 5D are diagrams schematically showing an image processing process in this processing. This process is realized by the CPU 201 executing a control program stored in advance in the memory 204 to control each part of the apparatus. Among these, various processes for the image are executed by the graphic processor 202, but at least a part thereof may be executed by the CPU 201.
  • the well plate WP carrying the spheroid as the imaging object in the well W together with the culture solution is carried into the imaging unit 1 and set in the holder 11 (step S101).
  • the imaging optical system 13 is positioned with respect to the well W to be imaged, and imaging by the imaging device 134 is performed (step S102). Thereby, the original image containing a spheroid is acquired.
  • the graphic processor 202 performs predetermined image processing on the original image thus obtained, and detects the area of the image object included in the original image (step S103).
  • a known technique can be applied to the extraction of the object in the original image. For example, it is possible to apply a method of binarizing an original image with an appropriate threshold value and dividing it into a background area and an object area.
  • the graphic processor 202 further extracts a spheroid-like object including a region corresponding to the spheroid from the detected image object (step S104).
  • the image object detected in step S103 may include a spheroid-only object, a spheroid-only object, and a spheroid-foreign object overlapping.
  • an object including a spheroid that is, an object composed only of a spheroid, and an object in which a spheroid and a foreign object overlap are extracted as a spheroid-like object.
  • a spheroid-like object having a circularity greater than or equal to a predetermined value can be used.
  • An object having an area larger than a predetermined circle of a predetermined size can be a spheroid-like object.
  • an object having an area equal to or larger than a predetermined value can be a spheroid-like object.
  • a condition for considering an object consisting only of a foreign object may be set, and an object that does not match the condition may be regarded as a spheroid-like object.
  • step S105 One is selected from the spheroid-like objects extracted in this way (step S105), and the minimum inclusion circle of the object is specified (step S106). If the ratio of the area of the minimum inclusion circle to the area of the object is equal to or greater than a predetermined threshold value greater than 1 (YES in step S107), the object is subjected to region separation processing in step S108. If the area ratio is smaller than the threshold value (NO in step S107), step S108 is skipped. This is because an object that does not include a foreign substance area is excluded from the target of the area separation process, and the image information is stored. It also helps shorten processing time.
  • the relationship between the object and the minimum inclusion circle is shown in FIG. 5A.
  • the minimum inclusion circle of the object OB is a circle C3 having the smallest diameter among circles including the entire object OB.
  • the diameter D3 of the minimum inclusion circle C3 is much larger than the diameter of the spheroid region Ra
  • the minimum inclusion circle C3 is a background region other than the object OB. Will contain a lot. Therefore, the area of the minimum inclusion circle C3 is larger than the area of the object OB.
  • the object OB does not include the foreign substance region Rb and consists only of the spheroid region Ra
  • the size of the minimum inclusion circle C3 is not significantly different from the size of the spheroid region Ra, and the area ratio approaches 1. It may be considered that the area ratio is less than 1 in principle.
  • the object OB does not include the foreign substance region Rb, and if the area of the minimum inclusion circle C3 is sufficiently larger than the area of the object OB, the object OB is It can be determined that the foreign object region Rb is included.
  • an appropriate threshold value for example, 1.1
  • an object OB having an area ratio equal to or greater than the threshold value is set as a region separation process target.
  • the object is also processed when the area ratio is equal to the threshold value.
  • whether such an object is processed is optional.
  • the region separation process (step S108) illustrated in FIG. 4 is performed on the object.
  • an approximate circle that approximates the spheroid region is specified for the object (step S201).
  • This approximate circle is a circle that approximates the spheroid region Ra included in the object OB by a circle, and is considered to have a size equivalent to the spheroid region Ra.
  • FIG. 5B shows an example of an approximate circle.
  • the spheroid region Ra has a shape close to a circle, and as a method of approximating it to a circle of the same size, for example, the outline of the spheroid region Ra without protruding from the object OB as shown by a circle C4 indicated by a broken line in the figure.
  • a circle having the maximum diameter among the circles that fit within the circle referred to herein as a “maximum inclusion circle”.
  • a circle having the same area as the area of the object OB including the spheroid region Ra and the foreign material region Rb (herein referred to as an “area equivalent circle”), such as a circle C5 indicated by a dotted line in the figure, is an approximate circle. You can also.
  • the diameter of the circle which is a structural element in the morphological process, is determined (step S202).
  • the morphological treatment in this case has a diameter of (1/6) to (1/2), more preferably (1/4) to (1/3) of the diameter of the spheroid region Ra. It has been found that good results are obtained when a circle is the structural element. However, since the spheroid region Ra and the foreign matter region Rb are undivided at this time, the size of the spheroid region Ra cannot be clearly specified. Therefore, the diameter of the approximate circle described above is regarded as the diameter of the spheroid region Ra. That is, the diameter of the circle as the structural element is determined in the range of (1/6) to (1/2) of the diameter of the approximate circle. Here, it is set to a value not less than (1/4) and not more than (1/3) of the diameter of the approximate circle.
  • the size of the structural element can be appropriately set according to the size of the spheroid and foreign matter, the purpose of processing, and the like. In any case, the size of the structural element is set according to the size of the spheroid region, thereby preventing the small spheroid from being erased by the erosion process.
  • the diameter of the circle as a structural element should be smaller than (1/2) of the diameter of the approximate circle. Is desirable.
  • the erosion process and the duration process are sequentially performed using the structural elements thus determined (steps S203 and S204).
  • the region Rd corresponding to the spheroid region Ra is reproduced in the image, while the foreign material region Rb is erased from the image.
  • the purpose of erasing the foreign object region Rb from the image is achieved at this point.
  • the purpose of extracting only the spheroid region Ra from the object OB is also achieved at this time.
  • the region separation processing of this embodiment it is aimed to clearly distinguish these regions while leaving the image information of both the spheroid region Ra and the foreign matter region Rb. That is, the spheroid region Ra identified by the processing up to step S204 is expanded by an appropriate size, for example, one pixel (step S205), and the result is subtracted from the original image (step S206).
  • the number of pixels to be expanded may be 2 pixels, for example.
  • the image of the spheroid region Ra and the foreign material region are added as shown in FIG. 5D by adding and synthesizing the image of the spheroid region Ra extracted by the duration processing (step S204) to the image from which the foreign material region Rb is extracted.
  • An image in which the image information of Rb is stored together and both areas are clearly separated is obtained (step S207). This image is an image to be obtained in this processing.
  • This composite image is stored in the image memory 203 (step S208). Instead of or in addition to this, an image of the spheroid region Ra (FIG. 2C) obtained by the process of step S204 and an image of the foreign substance region Rb (FIG. 5C) obtained by the process of step S206 are displayed in the image memory 203. It may be stored in.
  • step S109 The above processing (steps S105 to S108) is executed for all the extracted spheroid-like objects (step S109).
  • the diameter of the circle used as the structural element is individually set for each object. Therefore, a circle having a large diameter is applied as a structural element to an object including a large spheroid region, and a circle having a small diameter is applied to an object including a small spheroid region. Therefore, a small spheroid is not erased by the erosion process, and each object can be appropriately processed to efficiently distinguish the spheroid area and the foreign substance area.
  • the processed image is stored in the image memory 203 and is output in an appropriate form via the interface 205 (step S110).
  • the processed image is displayed on the display unit 22 and presented to the user.
  • the data is output to an external device or an external storage medium via the interface 205.
  • the first method performs the above-described morphological processing on the binarized original image to create a mask indicating the range of the spheroid region Ra and the foreign material region Rb, and applies this mask to the original multi-valued original image. By doing so, the image information of the spheroid region Ra and the foreign material region Rb is restored.
  • the second method is to restore the pixels in the region that is expanded when performing the duration process with the pixel values of the original image.
  • the imaging unit 1 and the image processing unit 2 function as an “image processing apparatus” of the present invention.
  • the imaging unit 1 functions as an “image acquisition unit” of the present invention
  • the holder 11 and the imaging unit 13 function as a “holding unit” and an “imaging unit” of the present invention, respectively.
  • the image processing unit 2, particularly the graphic processor 202 functions as the “image processing means” of the present invention.
  • the size of the structural element is determined by approximating the spheroid region Ra to a circle, but more generally, the spheroid region is preferably approximated by an ellipse.
  • the spheroid region may have a shape deviating from a circle due to distortion of the spheroids or overlapping of a plurality of spheroids on the image.
  • the diameter of the circle as the structural element is preferably set based on the minor axis of the approximate ellipse.
  • the minor axis of the ellipse In the departure from the circular shape of the spheroid region due to the above causes, it is unlikely that the minor axis of the ellipse is significantly different from the true size of the spheroid, while the major axis may greatly exceed the true size of the spheroid. It is possible.
  • determining the diameter of the circle based on the minor axis which is a smaller dimension, it is possible to prevent the spheroid from being erased from the object.
  • the image processing apparatus of the above embodiment includes the imaging unit 1 as “image acquisition means”, the present invention can also be applied to an apparatus that does not have an imaging function. That is, in the image processing unit 2 of the above embodiment, the interface 205 acquires an original image by receiving image data from the outside. In this sense, the interface 205 has a function as an “image acquisition unit”. Yes. As described above, the present invention functions effectively even in an aspect in which an image processing apparatus having no imaging unit receives original image data from an external apparatus or an external storage medium.
  • the present invention is implemented by the CPU 201 executing a control program stored in advance in the memory 204.
  • the image processing unit 2 in this embodiment is a general-purpose computer device. Can be used. Therefore, the present invention is provided to the user as a control program for causing the computer apparatus to execute the above-described image processing on the assumption that the image apparatus is read by such a computer apparatus, and in a mode in which the program is recorded on an appropriate recording medium. It is also possible to do so. Accordingly, for example, it is possible to add a new function to an imaging apparatus that is already in operation.
  • the present invention when there are a plurality of spheroid-like objects having different sizes, it is preferable that structural elements having different sizes are set for each.
  • the original image can include spheroids of various sizes.
  • the size of the spheroid-like object can be represented by the size of an approximate ellipse that approximates the spheroid-like object, for example.
  • the spheroid image is generally circular, but there is irregular shape at the periphery. For this reason, it is not always easy to specify the size strictly, and a simpler alternative method is required.
  • the size of the spheroid-like object may be the size of an ellipse that approximately represents the spheroid-like object. Thereby, the size of the structural element can be determined by a simple calculation.
  • an ellipse having the largest area among the ellipses included in the outline of the spheroid-like object may be set as the approximate ellipse.
  • an ellipse having the same area as the spheroid-like object may be an approximate ellipse.
  • Image processing algorithms corresponding to these methods for obtaining approximate ellipses have been put into practical use, and the processing can be simplified by applying them.
  • the diameter of the circle as the structural element is not less than (1/6) and not more than (1/2), more preferably not less than (1/4) and not more than (1/3) of the minor axis of the approximate ellipse. It may be configured as follows. According to the experiment by the present inventor, it has been found that good processing results can be obtained when the size of the structural element is within the above range. That is, it is possible to effectively remove the streak foreign object image while maintaining the shape reproducibility of the spheroid image.
  • the approximate ellipse may be an approximate circle that approximates a spheroid-like object. Since the spheroid is substantially circular, the size can be expressed with necessary and sufficient accuracy by circular approximation. In addition, the processing becomes simpler than when an approximate ellipse is used.
  • the present invention may include a step of detecting a non-spheroid region different from a spheroid among objects in the original image based on, for example, a difference image between the image after the dilation processing and the original image.
  • a difference image between the image after the dilation processing and the original image.
  • the method includes a step of identifying a minimum inclusion circle of the spheroid-like object detected in the original image, and the ratio of the area of the minimum inclusion circle of the spheroid-like object to the area of the spheroid-like object among the spheroid-like objects is A configuration in which the erosion process and the duration process are executed for a value larger than a predetermined value greater than 1 may be used. By doing so, spheroid-like objects that originally do not contain foreign objects and do not require processing can be excluded from processing, and the original image information can be stored.
  • image processing methods can be executed using a general-purpose computer. Therefore, the present invention may be provided to a user as a control program for causing a computer to execute the above processing.
  • the image acquisition means may include a holding unit that holds a carrier that carries spheroids, and an imaging unit that images the carrier and creates an original image. Good. Thereby, an original image can be acquired about various spheroids.
  • the present invention is suitable for use in imaging spheroids for the purpose of observation, analysis, etc., and can be used for various experiments in the fields of medicine, drug discovery, and biological science, for example.
  • Imaging unit image acquisition means
  • Image processing unit image processing means
  • Holder holding part
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PCT/JP2015/069894 2014-09-30 2015-07-10 画像処理方法、制御プログラム、記録媒体および画像処理装置 WO2016051913A1 (ja)

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WO2009142333A1 (ja) * 2008-05-22 2009-11-26 国立大学法人東京大学 画像処理方法、画像処理装置及び画像処理プログラム並びに記憶媒体
JP2010152456A (ja) * 2008-12-24 2010-07-08 Ricoh Co Ltd 画像処理装置及び画像処理方法、並びに当該画像処理方法を用いたプログラム
WO2012117647A1 (ja) * 2011-02-28 2012-09-07 三洋電機株式会社 観察プログラムおよび観察装置

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WO2009142333A1 (ja) * 2008-05-22 2009-11-26 国立大学法人東京大学 画像処理方法、画像処理装置及び画像処理プログラム並びに記憶媒体
JP2010152456A (ja) * 2008-12-24 2010-07-08 Ricoh Co Ltd 画像処理装置及び画像処理方法、並びに当該画像処理方法を用いたプログラム
WO2012117647A1 (ja) * 2011-02-28 2012-09-07 三洋電機株式会社 観察プログラムおよび観察装置

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