WO2004044639A1 - スライドガラス、カバーガラス及び病理診断システム - Google Patents
スライドガラス、カバーガラス及び病理診断システム Download PDFInfo
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- WO2004044639A1 WO2004044639A1 PCT/JP2003/013325 JP0313325W WO2004044639A1 WO 2004044639 A1 WO2004044639 A1 WO 2004044639A1 JP 0313325 W JP0313325 W JP 0313325W WO 2004044639 A1 WO2004044639 A1 WO 2004044639A1
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- color
- image
- micro
- slide glass
- measurement sample
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/34—Microscope slides, e.g. mounting specimens on microscope slides
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
Definitions
- the present invention relates to an image capturing system having at least a microscope for observing a measurement sample and an imaging device for capturing an image of the measurement sample obtained from the microscope. It concerns slide glass. Background art
- a network operation is performed in which an image of a measurement sample taken using a microscope is sent to, for example, a place different from the place where the image was taken, and the image is used there. It is becoming.
- Such network operations include, for example, operations such as telepathology in which pathologists observe and diagnose images of a measurement sample taken with a microscope at a remote location.
- operations such as telepathology in which pathologists observe and diagnose images of a measurement sample taken with a microscope at a remote location.
- the microscope diagnosis becomes more frequent, even if different observers observe the same measurement sample with different microscopes under different observation conditions and output using different image output media, It has become important to be able to share the same image information that can be objectively evaluated under a light source having a standard color tone.
- the slide glass is composed of a portion on which a sample (measurement sample) to be observed by a microscope device is mounted, a microscope image obtained by imaging the sample observed by the microscope device, and a microscope at the time of imaging. It has a portion where the device information indicating the state of various controlled parts of the device is recorded as an optically readable dot code pattern, and a portion where a reduced image of a microscope image is recorded as an index image.
- This slide glass is intended to allow the collective management of specimens and information about the specimens.
- the above color chart is an industry standard, this color chart is used as a common reference to allow devices (eg, hardware such as color cameras, displays, and printers) within a specific manufacturer to be used. In addition to this, it is possible to objectively evaluate the color of the image of the measurement sample taken between devices manufactured by arbitrary manufacturers, as well as between devices manufactured by arbitrary manufacturers. However, little has been studied so far on the color evaluation of images under a microscope, and even on imaging systems including microscopes. [0107] In addition, as in the conventional practice, the observer can change the color appropriately according to his / her own sense of color (sensitivity), such as changing the color of the light source, or the observation technique. As important.
- the problem regarding the item (a) is caused by the difference in spectral sensitivity characteristics between the color cameras used. This challenge is more pronounced between manufacturers with different manufacturers.
- the ideal spectral sensitivity characteristic of a camera is determined based on the color system for display. For example, taking the s RGB color coordinate system, which has recently become a standard, as an example, its ideal sensitivity characteristics are the chromaticity coordinates of each RGB color in the s RGB color coordinate system and those shown in Figure 1. It is derived from the color matching functions of the XYZ color system (X ( ⁇ ), y (e), z ( ⁇ )).
- the ideal spectral sensitivity characteristics of this color system usually have negative sensitivity characteristics (for example, see Fig. 2).
- any other coordinate system eg, PAL, NTSC (1953), CIERGB, sony's trinitron RGB coordinate system
- each manufacturing manufacturer performs a matrix operation based on the camera sensitivity characteristics (raw data) shown in FIG. 3, for example, as shown in FIG.
- Such camera sensitivity characteristics are created.
- An ideal spectral characteristic having no negative sensitivity and all positive values is a color matching function of the XYZ color system shown in FIG. From now on, any ideal imaging characteristics One-to-one conversion is possible. However, for example, it is difficult to create this ideal imaging characteristic with a prism of a three-plate camera. It is common to create from the sensitivity characteristics (raw data) of the data.
- the basic spectral portion which is the sensitivity characteristic (raw data) of the camera shown in FIG. 3, and a matrix calculation for correcting the same (formerly processed by an analog circuit)
- the final spectral characteristic which is the sensitivity characteristic (correction data) of the camera shown in Fig. 4, obtained by the above is the part where the characteristics of each manufacturer appear, and the saturation is emphasized. Or redness is emphasized. Therefore, in the case of color cameras, the spectral sensitivity characteristics differ from manufacturer to manufacturer, and the final spectral sensitivity characteristics have not been clarified.
- the issues regarding the above item (b) are as follows. That is, when the measurement sample is photographed through a microscope, the color temperature (color) of the light source is appropriately adjusted to highlight the characteristic difference between the measurement samples, and the color balance is adjusted as necessary. Is adjusted, and the image of the best view of the observer is acquired. For this reason, the data values relating to the color of the image of the first measurement sample are often ignored, and absolute values are lost.
- the slide glass according to the present invention includes a glass plate and one or more micro color filter groups provided on one surface of the glass plate.
- each of the one or more micro color filter groups is composed of two or more color reference micro color filters for sampling a color used as a reference for color evaluation and color correction of an image.
- two or more color reference micro color filters belonging to the same group among the one or more micro color filter groups are arranged on the surface of the glass plate so as not to overlap each other, and different reference colors from each other.
- the color reference micro color filter includes a color reference micro gray filter having an achromatic color plotted on an achromatic axis.
- imaging device includes components other than a camera, such as a digital no-slide (vertical reslide), a scanner, an imaging unit in a Virtual Microscope, and other components for imaging a sample. Is included.
- color in “color evaluation and color correction of images” and “color” in “comparative reference color” are created from the spectral distribution of the measurement sample.
- the spectral characteristics (or any R, G, B spectral characteristics) such as color matching functions (X (e), y ( ⁇ ), ⁇ ( ⁇ )), etc., by the light source spectral distribution or radiation spectrum itself It is a value obtained by the above, or its modified value (L * a * b *).
- the typical name that the value means, for example, red (red), yellow (yel low), blue (blue), etc., or a special technical name (for example, 1.6YR6 on the Munsell color chart value) 3/3. 9 etc.). This is also consistent with human visual perception.
- color information which is a comparison standard usable for color evaluation and color correction of an image, indicates information on a color in consideration of the information of the above-described spectral characteristics.
- the "spectral distribution of the measurement sample” indicates the spectral distribution of the measurement sample itself.
- a spectral reflectance, a spectral transmittance, a spectral emissivity, and the like obtained by a usual spectroscopic measurement method.
- the slide glass according to the present invention (hereinafter referred to as “slide glass with a color reference microfilter” as required) is provided with the color characteristics (spectral characteristics) of the color reference microcolor filter. ) Can be used as a comparison standard to objectively evaluate the color characteristics of the image of the measurement sample photographed with a microscope.
- an image output unit After adjusting the color of the final image output in step 2 so that it matches the visual image obtained by microscopy, adjusting the color to the color desired by the user, and adjusting the color to the color desired by the user, the first Correction to restore the color characteristics of the original image, etc.
- images taken by different imaging devices are output by different image output means (output images on display monitors, paper using printers, etc.) It is also possible to reliably evaluate and calibrate the output image on the recording medium.
- the slide glass with the color reference microfilter according to the present invention, various images are added to the original image of the measurement sample before the final image output by the image output means is obtained.
- the measurement conditions of the color characteristics (spectral characteristics) of the color reference micro-color filter eg, characteristics such as the color temperature of the light source during shooting, lens system characteristics, sensitivity characteristics, and internal
- the degree of color value change by software for each processing step performed how the color characteristics (spectral characteristics) of the color reference microcolor filter changed for each processing It is easy to grasp.
- the above-described quantitative determination is performed by using a spectral imaging sensor (for example, a spectral imaging sensor described in Japanese Patent No. 2713838) instead of an RGB color camera. You may go as. Thereby, for example, the concentration of the stained measurement sample can be measured.
- a spectral imaging sensor for example, a spectral imaging sensor described in Japanese Patent No. 2713838
- the concentration of the stained measurement sample can be measured.
- the color reference micro color filter for example, a filter containing a reference color material at a reference density may be prepared. Based on this, the unknown concentration of a specific substance in the measurement sample can be determined under a microscope.
- the slide glass (slide glass with a color reference microfilter) according to the present invention provides information on an invisible color of an image of a measurement sample. May be acquired in the same manner as in the case of handling the information of the color included in the visible region described above.
- the invisible color image of the measurement sample contains extremely useful information on the physical or chemical state of the subject that cannot be recognized by the human eye, the slide glass is used.
- the acquired image information may include information on an invisible color image.
- the “invisible color” refers to the visible light (light having a wavelength in the range of 380 to 780 nm) contained in the radiation emitted from the image of the measurement sample. Colors based on non-visible light (e.g., ultraviolet light having a wavelength in the range of 200 to 380 nm or light having a wavelength in the range of 780 nm to 20 ⁇ m near-infrared light) Show.
- non-visible light e.g., ultraviolet light having a wavelength in the range of 200 to 380 nm or light having a wavelength in the range of 780 nm to 20 ⁇ m near-infrared light
- the color value (invisible) can be obtained by the same method as that for handling the spectral distribution of visible light. Further, another color value can be obtained by multiplying the color value by an appropriate integration function. Further, a representative name represented by the color value obtained in this way is defined as a color name. In this specification, color names determined based on these invisible lights are collectively referred to as
- the "reference color” is ideally a so-called unique color, which is generally said to be a color that is not mixed among hues such as red, green, yellow, and blue. It is only necessary that the hues between colors are separated at a uniform rate, and it is not necessary to define exact hue values.
- the reference color is effective in two or more colors, preferably three or more colors. If the hue angle between each reference color is about 120 degrees, four colors are about 90 degrees, six colors are about 60 degrees, and 12 colors are about 30 degrees, etc. It does not matter which hue is used as a reference as long as the color is set so that the interval is as uniform as possible.
- an image acquisition system that uses the slide glass with the micro filter for color reference In, the colors that can be displayed are the minimum requirements.
- the color developed by the dye is the reference color.
- at least two colors can be used as reference colors, and color value confirmation and color adjustment are performed within the range of colors that can be formed between at least two colors.
- the color of the achromatic axis is also matched (that is, the color without any sample is matched with white (gray)), so if this is included, three colors are used.
- the invisible color criterion (5) to be described later is basically the same as the visible light region. That is, the hue of the invisible color value obtained by multiplying the measured spectral value of each part by the sensitivity function of an imaging device, such as a camera, at the same rate as in the criteria of (1) and (2), It is also possible to select such a sample, measure it, and use the color set as the reference. Also, in the same way as the reference in (3), the sample dyed with the reference dye and the spectral value for each site The invisible color value itself obtained by multiplying the sensitivity function of the imaging device may be used as the reference value.
- one micro color filter group for example, a microscope observation color pallet unit 3a, 3b, 3c, 3d
- the number of color reference micro color filters included in the same micro color filter group is two or more
- the two or more color reference micro color filters belonging to the same micro color filter group are different reference colors.
- one microcolor filter group includes information of at least two or more reference colors of all hues.
- a group of microcolor filters (for example, a color microscope for microscopic observation in FIG. 5 described later, let portions 3a, 3b) , 3c, 3d) 1 Provided for each objective lens provided in the microscope.
- Two or more color reference microcolor filters belonging to the same group among one or more micro color filter groups have respective sizes. Is preferably adjusted to a range that can be captured in the field of view obtained when the corresponding objective lens is used.
- the user can place a micro color filter group having a size matching the magnification of the objective lens within the field of view.
- the color can be captured, and the color used as a reference for color evaluation and color correction of the image of the measurement sample can be accurately and easily grasped.
- two or more color reference microcolor filters belonging to any one of the one or more microcolor filter groups are visible. (See large area color palette section 2 in FIG. 5 described below).
- the user can easily understand what kind of color the micro color filter group using the color reference micro filter is used without using a microscope. .
- the user can measure the spectral characteristics of the color reference microfilter without using a microscope.
- two or more color reference microcolor filters belonging to the same group among one or more microcolor filter groups have the same size. It is preferred to have This makes it easy to arrange the micro color filters for each color reference on the surface of the glass plate compactly and densely, and it is easy to configure one micro color filter group compactly. .
- the one or more microcolor filter groups have an intermediate chroma color having a chroma difference of equal rate with respect to the reference color.
- One or more color reference micro color filters may be further provided. This makes it possible to more accurately perform color evaluation and color correction of an image.
- the slide glass with the color reference microfilter according to the present invention is applied to the slide glass.
- the one or more micro color filter groups may further include one or more color reference micro color filters having an achromatic color plotted on an achromatic axis. This makes it possible to more accurately evaluate and correct the color of the image (particularly the color of the neutral axis).
- each of the reference colors is preferably red, green, blue, cyan, magenta, or yellow.
- two or more color reference microcolor filters belonging to the same group among one or more microcolor filter groups each have a rectangular shape. And are arranged at predetermined intervals so as not to contact each other.
- the slide glass with the color reference micro filter further includes an annular black micro filter arranged so as to surround the entire peripheral portion of each color reference micro color filter without a gap.
- two or more color reference microcolor filters belonging to the same group among one or more microcolor filter groups each have a rectangular shape. And may be juxtaposed in a dense state without any gaps.
- the micro color filters for each color reference can be juxtaposed compactly and juxtaposed on the surface of the glass plate more reliably and easily, and one micro color filter group can be compactly configured. Can be done more reliably and more easily.
- the slide glass with the color reference microfilter further includes an annular black microfilter arranged so as to surround the entire periphery of each microcolor filter group without a gap. I'm sorry. As a result, it is possible to sufficiently reduce the amount of sneaking light (for example, background light to slide glass during observation) from inside the microscope other than the part being observed.
- a measurement sample mounting portion for mounting the measurement sample is further provided on the surface of the glass plate. This allows the user to obtain the image, evaluate the color of the image, and correct the color at the same time.
- the slide glass with color reference microfilters according to the present invention has a structure in which two or more color reference microcolor filters are printed on a glass plate.
- the micro color filter may be printed on a cover glass which is provided on the glass plate and which corresponds to the measurement sample mounting portion. That is, on one surface of the cover glass (cover glass according to the present invention), at least one color reference for sampling a color used as a reference for color evaluation and color correction of the image is provided.
- a micro color filter is arranged.
- the color reference micro color filters are adjusted so that each size is within a range that can be captured in a field of view obtained by using at least one of the objective lenses provided in the microscope. You. ,
- the color reference micro color filters are preferably arranged asymmetrically with respect to the center of gravity of the power bar glass. In this way, the asymmetric arrangement of the color reference micro color filters makes it possible to clearly distinguish the upper surface and the rear surface of the cover glass.
- the cover glass may be provided with an identification mark for distinguishing the top surface from the back surface.
- the slide glass having the above-described structure can be applied to a pathological diagnosis system.
- the pathological diagnosis system performs a pathological diagnosis using an image of a measurement sample placed on the slide glass.
- the pathological diagnosis system converts an image of a measurement sample placed on a slide glass, taken through the cover glass, into an image. A pathological diagnosis is made by utilizing this.
- a pathological diagnosis system includes at least an image collection system, an image converter, and a monitor, and converts image data of a measurement sample collected by the image collection system into Remote pathological diagnosis using wired or wireless communication lines (electronic communication lines such as LAN and Internet, satellite lines, telephone lines, etc.) is also possible.
- wired or wireless communication lines electronic communication lines such as LAN and Internet, satellite lines, telephone lines, etc.
- the image collection system is a microscope for observing a measurement sample, and an imaging device for collecting an image of the measurement sample obtained from the microscope. (Virtual slide), scanner, Virtual
- the image converter captures the color information of the color reference micro color filter taken by the image capturing system directly or through a communication line, and displays an image to be displayed based on the color reference micro color filter. Adjust the color of. Further, the monitor displays an image of the color adjusted by the image converter.
- FIG. 1 is a graph showing the color matching functions (XU), y (e), and ⁇ ( ⁇ ) of the XYZ color system.
- FIG. 2 is a graph showing an example of the ideal sensitivity characteristic of the s R G ⁇ color coordinate system.
- FIG. 3 is a graph showing an example of sensitivity characteristics (raw data) of a camera used for microscopy.
- FIG. 4 is a graph showing an example of sensitivity characteristics (correction data) of a camera used for microscopic photographing.
- FIG. 5 is a front view schematically showing a basic configuration of a slide glass (a slide glass with a color reference microfilter) according to a first embodiment of the present invention.
- Fig. 6 shows the chromaticities of the main reference colors assigned to any of the microphone opening filters MF1 provided on the slide glass (slide glass with color reference micro-finolators) shown in Fig. 5. It is a graph showing an example of a value on coordinates (a * b * space).
- Fig. 7 shows the filter characteristics of red (reference color).
- rl is red (high chroma color)
- r2 is red (medium chroma color)
- r3 is red (low chroma color).
- Fig. 8 shows the filter characteristics of yellow (reference color), in which yl is yellow (high chroma color), y2 is yellow (medium chroma color), and y3 is yellow (low chroma color).
- FIG. 9 shows the filter characteristics of green (reference color), where gl is green (high chroma color), g 2 is green (medium chroma color), and g 3 is green (low chroma color).
- Fig. 10 shows the filter characteristics of cyan (reference color).
- cy1 is cyan (high chroma color)
- cy2 is cyan (medium chroma color)
- cy3 is cyan. (Low saturation color).
- FIG. 11 shows the filter characteristics of blue (reference color).
- b 11 is blue (high chroma color)
- b 12 is blue (medium chroma color)
- b 13 Indicates blue (low saturation color).
- FIG. 12 shows the purple (reference color) filter characteristics.
- pr 1 is purple (high chroma color)
- Pr 2 is purple (medium chroma color)
- pr 3 is purple. (Low-saturation color).
- FIG. 13A is a front view schematically showing a basic configuration of a slide glass (slide glass with a color reference microfilter) according to the second embodiment of the present invention
- FIG. FIG. 2 is a front view showing a cover glass.
- FIG. 14 is an enlarged view of a region R1 in the measurement sample mounting portion shown in FIG. 13A.
- FIG. 15 is an enlarged view of a region R3 in the region R1 shown in FIG.
- FIG. 16A is a front view schematically showing a basic configuration of the slide glass (slide glass with color reference micro finoleta) according to the third embodiment of the present invention
- FIG. FIG. 2 is a front view showing a cover glass.
- Fig. 17 shows a display example when an image of a measurement sample taken using the slide glass (slide glass with a color reference microphone opening filter) shown in Fig. 16 A is output.
- FIG. 17 shows a display example when an image of a measurement sample taken using the slide glass (slide glass with a color reference microphone opening filter) shown in Fig. 16 A is output.
- FIG. 18 shows another display example in which an image of a measurement sample taken using the slide glass (slide glass with a color reference microphone opening filter) shown in FIG. 16A is output.
- FIG. 18 shows another display example in which an image of a measurement sample taken using the slide glass (slide glass with a color reference microphone opening filter) shown in FIG. 16A is output.
- FIG. 19 shows the color characteristics of each micro color filter MF 1 (spectral characteristics).
- FIG. 6 is a diagram for explaining an example of an L * a * b * color system used for setting of a color value.
- Fig. 20 shows the color values obtained when the sample was stained by the hematoxylin and eosin (HE) staining method expressed as a * and b * values in the L * a * b * color system. It is a graph obtained when doing.
- HE hematoxylin and eosin
- FIG. 21 is a graph obtained when the color values obtained when the measurement sample is stained by the Azan dye method are expressed as a * and b * values in the L * a * b * color system. is there.
- Fig. 22 is obtained when the color values obtained when the measurement sample is stained by the Watanabe silver plating method are expressed as a * and b * values in the L'a * b * color system. It is a graph.
- FIG. 23 shows the results obtained when the color values obtained when the measurement sample is dyed by the Elastica and Wang Gieson staining method are expressed as the a * value and the b * value in the L * a * b * color system. It is a graph.
- Fig. 24 shows the results obtained when the color values obtained when the measurement sample is stained by the Masson trichrome staining method (MT) are expressed as a * and b * values in the L'a * color system. It is a graph.
- MT Masson trichrome staining method
- FIG. 25 is a diagram for explaining an example of image processing using the slide glass (slide glass with a color reference microfilter) according to the present invention.
- FIG. 26 is a diagram for explaining an example of image processing using the slide glass (slide glass with a color reference microfilter) according to the present invention.
- Figs. 27A and 27B are front views schematically showing the basic structure of the cover glass according to the first and second embodiments of the present invention.
- FIG. 28 is a diagram showing a schematic system configuration as a pathological diagnosis system according to the present invention, particularly when performing remote pathological diagnosis.
- Fig. 29 is a flowchart for describing preprocessing up to the start of remote pathological diagnosis in the pathological diagnostic system shown in Fig. 28.
- Fig. 5 is a front view schematically showing a basic configuration of the slide glass according to the first embodiment of the present invention (hereinafter, referred to as "slide glass with color reference microphone opening filter").
- the slide glass 10 with a color reference microfilter includes a glass plate 1 (for example, a size of 26 mm ⁇ 76 mm) and an upper surface of the glass plate 1. It consists of a large-area color palette section 2 formed on F1, four color palette sections 3a, 3b, 3c, 3d for microscopic observation, and a name label 4.
- the large-area color pallet unit 2 has at least two micro color filters (transmitters) MF 1 having different colors (22 in FIG. 5). ) Force to be inserted ⁇ Force to be stuck ⁇ Or micro color filter group placement area 22 juxtaposed in printed state and black micro filter MF2 surrounding the outer edge of this micro color filter group placement area 22 And the black microfilter disposition area 24 that has been set.
- Each micro color filter MF 1 in the micro color filter group arrangement area 22 has a substantially rectangular shape, and its size is, for example, about 4 mm X 3 mm.
- the large-area color palette section 2 is provided for measuring the spectral characteristics of each micro color filter MF1 without using a microscope.
- the black microfilter MF2 in the black microfilter disposition area 24 is a device in which the sneaking light (for example, the background light to the slide glass during observation) from a portion other than the observation site is a microscope. It is provided to sufficiently reduce ingress.
- the black microfilter disposition area 24 can be formed on the upper surface F1 of the glass plate 1 by a known technique such as a black resist. In the case of measurement including the external sneaking light, the configuration may be such that the black microfilter arrangement area 24 is omitted.
- the color palette sections 3a, 3b, 3c, and 3d (micro color filter groups) for microscopic observation are similar in size to the large-area color palette section 2 with a certain size. It has the same configuration as the large area color pallet 2 except that it is reduced in size.
- the size of the color palette sections 3a, 3b, 3c, and 3d for microscopic observation is determined by the objective lenses (for example, 10x, 20x, and 40x) provided at different magnifications provided in the microscope used. , An objective lens having a different magnification such as 100 times). Therefore, even when changing the area of the visual field capture simultaneously replaced with a different magnification microscope objective lens, any one of the observer force microscope observation error pallet unit 3 a, 3 b, 3 c , 3 d Can always be captured in the field of view.
- a notch 1a is provided in the outer extension of the glass plate 1 (in Fig. 5, one of the four corners of the rectangular upper surface F1). Thereby, the user can easily distinguish the upper surface F1 and the rear surface of the glass plate 1 from each other.
- the name label 4 is provided for writing notes such as the name of the measurement sample and observation conditions of the measurement sample.
- the micro color filter MF 1 and the black micro filter MF 2 of the large area color palette section 2 and the color palette sections 3a, 3b, 3c, 3d for microscopic observation are respectively Printed under the same conditions (same process, same time). Therefore, since the various characteristics of the printing parts are equal, they have the same spectral characteristics.
- the micro color filter MF1 provided in each of the large area color palette section 2 and the color palette sections 3a, 3b, 3c, 3d for microscopic observation will be described.
- Each micro color filter MF 1 has a transmission material (e.g., 6 colors of the main reference colors of red, green, blue, cyan, magenta, and yellow) selected to have a uniform hue. Color material).
- the reference color may be a little more fine, and may be 8 or more colors, 10 or more colors, or 12 or more colors.
- a filter composed of several levels of tint of these reference colors (a color having a chromaticity characteristic whose saturation is gradually reduced from the main reference color) may be further arranged.
- achromatic (transparent, gray, black) filters may be arranged. Three micro color filters with only the three primary colors (red, green, and blue) can be easily used.
- the large area color pallet section 2 and the color pallet sections 3a, 3b, 3c, and 3d for microscopic observation can be obtained using only the MF1. It may be composed.
- the reference color is minimized. obtain.
- a filter array having a different scatterer may be used. This allows it to be used as a scatterer reference for microscopes.
- the "primary reference color” is a color that falls within the chromaticity coordinates that can be expressed by a normal display system.
- the chromaticity coordinates (a * b * space) is a color having a spectral transmittance characteristic designed to match.
- the “spectral transmittance characteristics” are, for example, the transmittance characteristics of filters as shown in FIGS. 7 to 12.
- Fig. 7 shows the filter characteristics of red (reference color).
- rl is red (high saturation color)
- r2 is red (medium saturation color)
- r3 is red (low saturation).
- saturated color e.g. 8
- y1 is yellow (high chroma color)
- y2 Indicates yellow (medium chroma color)
- y3 indicates yellow (low chroma color).
- Fig. 9 shows the filter characteristics of green (standard color).
- g1 is green (high chroma color)
- g2 is green (medium chroma color)
- g3 is green (low chroma color).
- Fig. 10 shows the filter characteristics of cyan (reference color).
- cyl is cyan (high chroma color)
- cy2 is cyan (medium chroma color)
- cy3 is cyan (low chroma).
- b11 is blue (high chroma color)
- b12 is blue (medium chroma color)
- b13 is blue (low chroma color).
- Fig. 12 shows the purple (reference color) filter characteristics.
- pr 1 is purple (high chroma color)
- Pr 2 is purple (medium chroma color)
- pr 3 is purple (low chroma color).
- a microfilter of a main reference color and a color having chromaticity characteristics in which the saturation is gradually reduced from the main reference color may be, for example, adjustment of the thickness of the microfilter. It can be formed by a technique such as reducing the concentration of a color material (each color material).
- the slide glass 10 with the color reference microfilter is used as follows. That is, a slide glass 10 with a color reference microfilter is prepared, and a slide glass for mounting a measurement sample is prepared. Then, they are interchanged, and the color reference and the measurement sample are separately photographed.
- FIG. 13A is a front view schematically showing a basic configuration of the slide glass (slide glass with color-reference microfilter) according to the second embodiment of the present invention
- FIG. FIG. 2 is a front view showing a cover glass
- FIG. 14 is an enlarged view of a region R1 in the measurement sample mounting portion 5 shown in FIG. 13A.
- the region R1 shown in FIGS. 13A and 14 shows, for example, the region of the microscope field of view when an objective lens with a magnification of 10 times is used.
- R 2 in FIG. 14 indicates the area of the field of view of the collected image.
- FIG. 15 is an enlarged view of a region R3 in the region R1 shown in FIG.
- a region R3 shown in FIGS. 14 and 15 indicates, for example, a region of a microscope visual field (or a collected image region) when an objective lens having a magnification of 100 times is used.
- the slide glass 1 OA with the color reference microfilter according to the second embodiment is a glass plate 1 (for example, a size of 26 mm X 76 mm), a large-area color palette portion 2 formed on the upper surface F1 of the glass plate 1, a measurement sample mounting portion 5, a bar code 6, and a memo space 7.
- the glass plate 1 and the large-area color pallet unit 2 are the same as those of the slide glass 10 with a color reference microphone opening filter according to the first embodiment described above.
- the measurement sample mounting portion 5 is a portion on which the measurement sample is mounted, and the cover glass 20 of Fig. 13B is abutted with the measurement sample interposed therebetween.
- a plurality of color pallets 3 e for microscope observation are provided on the measurement sample mounting portion 5. Mutual They are juxtaposed at predetermined intervals so that they do not overlap.
- Each of these color pallet sections 3e for microscopic observation has the same configuration as the large area force pallet section 2 except that it is reduced at a certain similar ratio to the large area color pallet section 2.
- the size of each microscopic observation color palette section 3 e is, for example, 50 ⁇ m ⁇ 40 im.
- the barcode 6 shown in Fig. 13A indicates the color information and the Z of the image of the measurement sample obtained by microscopic imaging or the measurement conditions at the time of image acquisition (for example, the type of light source, the spectrum of the camera). It is prepared to record additional information such as sensitivity.
- the bar code 6 may be formed of, for example, a black color material, or may be formed of a color material other than black.
- the memo column 7 shown in FIG. 13A is provided for the same purpose as the bar code 6. This memo field 7 may be formed, for example, by applying a magnetic material.
- the storage form of the original information that is, the preparation itself
- the color of the image of the measurement sample obtained by the microscopic photographing is also continued.
- additional information can be added to the slide (slide glass + measurement sample) itself.
- the additional information includes, for example, the measurement date and time, the sample name, the imaging conditions (the imaging light source, the sample lens, the camera used, and the like).
- the slide glass 1 OA with the color reference microfilter is a color reference microfilter having a size corresponding to the objective lens used in the area of the measurement sample mounting section 5 on which the measurement sample is mounted.
- the power of the color filter being fitted and pasted Alternatively, it has a printed configuration, and when the area of the measurement sample mounting portion 5 is photographed, the reference micro color filter MF1 can be confirmed together with the measurement sample S1 in the field of view.
- the micro color filter for color reference is embedded or printed on the upper surface F 1 of the glass plate 1.
- the force embedded on the cover glass 20 that is in contact with the mounting portion 5 may be printed (the cover glass according to the present invention).
- FIG. 16A is a front view schematically showing a basic configuration of a slide glass (slide glass with a color reference microfilter) according to a third embodiment of the present invention
- FIG. FIG. 2 is a front view showing a cover glass.
- the slide glass 10 B with the color reference microfilter according to the third embodiment is a glass plate 1 (for example, a size of 26 mm X 7 6 mm), a large-area color pallet 2 formed on the upper surface F 1 of the glass plate 1, a color pallet 3a, 3b, 3c, 3d for microscope observation, and a measurement sample mounting part. 5a, a positioning reference mark 8, a bar code 6, and a memo box 7.
- the cover glass 20 in FIG. 16B is brought into contact with the glass plate 1 with the measurement sample sandwiched therebetween.
- the glass plate 1, the large-area color pallet unit 2, and the color pallet units 3a, 3b, 3c, and 3d for microscopic observation are the color reference microphone opening filters according to the first embodiment described above. It is the same as the attached slide glass 10. Also, barcode 6 and memo field 7 is the same as the slide glass with color reference microfilter 10A according to the second embodiment described above.
- the measurement sample mounting portion 5a shown in FIG. 16A is the same as the above-described color reference micro device according to the second embodiment except that the color pallet portion 3e for microscope observation is not provided. It has the same configuration as the measurement sample mounting part 5 of the slide glass with filter 1 OA.
- the positioning fiducial mark 8 is used when the color palette section 3a, 3b, 3c, or 3d for microscopic observation is photographed. , Corresponding to the X-Y stage 311 in Fig. 29).
- the measurement sample S1 on the slide glass is automatically measured, and it is considered that the inspection and the like will also be automated.
- the slide glass 10B with a color reference microfilter shown in Fig. 16A is intended for use in such a situation.
- the measurement sample using the slide glass 1 OB with a color reference microfilter can be photographed, for example, as follows. That is, from a case in which a plurality of slide glasses with color reference microfilters 10B are stored, one slide glass with color reference microfilters 10B is taken out one by one and the mechanism is set using a mechanism that can be set on a microscope table. Slide glass 10B is set on the microscope table. After the slide glass 1 OB is set, the microscope stage moves so that the positioning reference mark 8 matches the predetermined coordinate position in the microscope field of view. After the positioning reference mark 8 is aligned with the coordinate position, the color pallet 3a for microscope observation is obtained by using an objective lens having a different magnification such as 10 times, 20 times, 40 times, or 100 times.
- the image of the measurement sample obtained from the microscope as described above is taken into an image collection system including a combi- ter and the like, and is automatically inspected according to the automatic inspection software provided for the system.
- the image of the measurement sample and the color palette for microscopy 3a is taken into an image collection system including a combi- ter and the like, and is automatically inspected according to the automatic inspection software provided for the system.
- An image of any of 3b, 3c, and 3d is synthesized. Then, the composite image is output from an image output unit such as a monitor / printer.
- the image of the measurement sample obtained from the microscope is transferred to another terminal via the existing communication line, whether wired, wireless, such as LAN, Internet, telephone line, satellite line, etc. (Telepathological diagnosis system). Also, this composite image may include the information described in the bar code 6 and the memo or memo field 7. As a result, the conditions at the time of photographing or information obtained from images obtained by photographing (information obtained by evaluating images such as diagnostic results) can be left as a more reliable record.
- FIG. 17 is an example of a composite image when the image is output to a monitor display, for example. Any of the microscopic observation color palette sections 3a, 3b, 3c, and 3d that match the magnification of the objective lens and the information described in the bar code 6 are displayed together.
- a composite image as shown in Fig. 18 may be output.
- the composite image shown in FIG. 18 is an example of a case where the composite image shown in FIG. 17 is printed on an information recording medium such as paper using a printer, for example.
- the marginal portion 12 of the information recording medium outside (outer edge) of the collected image area 11 in FIG. Any of 3b, 3c and 3d and the information described in bar code 6 may be displayed.
- the setting method of the color characteristic value (spectral characteristic value) of each micro color filter MF 1 provided in the slide glass with each color reference micro filter according to the first to third embodiments described above. Will be described.
- the setting method the following setting method (1) is basically adopted depending on the information to be obtained from the image of the measurement sample.
- the setting methods (2) to (5) shown below are used. Is also good.
- each micro color filter MF1 As a color of each micro color filter MF1 , Typically 6 colors (colors selected to be equi-equivalent in hue, for example, 6 of the main reference colors red, green, blue, cyan, magenta, and yellow) or 12 colors A color having a different saturation is set as a reference. (2) As shown in Fig. 19, the color of each micro color filter MF1 has a step in the L * value (brightness) in the same a * b * color system to Covers the range occupied and consists of representative colors.
- the lightness axis is divided into seven levels, seven achromatic colors are set at the center, and 108 chromatic colors are configured.
- color standards are set for all three dimensions in this way, the amount of deviation of the sample color can be accurately estimated from the amount of deviation of the reference color, and this is used to achieve good color reproduction on a display and good color on a printer. The reproduction can be adjusted.
- FIG. 20 shows that a biological tissue sample as a measurement sample is stained by a hematoxylin and eosin (HE) staining method, which is a typical staining method for obtaining a reference color.
- HE hematoxylin and eosin
- This is a graph in which the color values are expressed as a * and b * values in the L * a * b * color system.
- he bl is the chromaticity point of the violet region of the nucleus etc. stained with hematoxin
- he pi is the chromaticity point of the reddish purple region such as the cytoplasm stained with eosin. Show.
- Fig. 21 shows the criteria for staining the measurement sample by selecting the Azan staining method as the staining method for connective tissue, in contrast to the hematoxylin and eosin (HE) staining shown in Fig. 20.
- 5 is a graph in which color values for colors are expressed as a * values and b * values in the L * a * b * color system.
- azan bl is the chromaticity point of the region stained in blue, which indicates the reticulo-reticular basal lamina, centered on the dark blue of the collagen fibers
- azan rl is stained red, mainly in the nucleus.
- the chromaticity point of the region, azanyl indicates the chromaticity point of the region dyed yellow otherwise.
- Figure 22 shows that the color value for the reference color when the measurement sample is dyed by the so-called Watanabe silver method (Reticulin silver inpregnation) is a in the L * a * b * color system. It is a graph expressed as a direct and b * value.
- ag liv bl is the chromaticity point of the dark and low-brightness blue (almost black) area centered on the nucleus and reticulum fibers
- ag liv pi is the other The chromaticity points of the purple dyed area are shown.
- FIG. 23 shows that the color value for the reference color when the measurement sample was dyed by the Elastica and Wang Giesson staining method (E 1 elasticavan Giessonstan (EVG)) was a * in the a * b * color system.
- 4 is a graph expressed as values and b * values.
- evgrl is the chromaticity point of the red area centered on collagen fibers
- evgbl is the dark and low-brightness blue area (almost black color area) centered on the nuclei and elastic fibers.
- the chromaticity point of the separated area, evgyl indicates the chromaticity point of the area dyed yellow, centering on red blood cells and cells, respectively.
- Figure 24 shows the color values for the reference color when the measurement sample was stained by Masson's trichrome staining (MT) expressed as a * value and value in the La * b * color system. It is the graph which was done.
- MT Masson's trichrome staining
- mt rl is the chromaticity point of the region stained red, such as cytoplasm or eosinophilic cell secretory granules
- mt bl is the chromaticity point of the region stained blue, such as other collagen fibers. Shown respectively.
- polysaccharide staining represented by periodic acid Schiff (PAS) staining, best carmine staining, Chiel'Nelsen staining, and Gram staining Staining of pathogens in tissues represented by the method, staining of blood cells in tissues, staining of kidneys, amyloid staining, staining of endocrine granules, staining of in vivo pigments, staining of inorganic substances in tissues, fat staining, central nervous system It is also possible to select a reference color that is close to the color developed by each staining method for tissue staining, nucleic acid staining, and enzyme histology staining. Also, in order to match the colors dyed by the respective dyeing methods, a color-matched color (metameric match color) using another color material may be used as a color reference.
- PIS periodic acid Schiff
- this color reference setting method (3) differs from the setting methods (1) and (2).
- the setting methods (1) and (2) three or more reference colors are set so that the color gamut to be used is approximately covered. Is a reference skeleton, and the measured value is determined based on the assumption that the measured value will be positioned (plotted) in the plane or in the solid, or outside the plane or the solid that has been extrapolated. It is. In other words, based on the line (skeleton) connecting the reference colors, it is important to determine where the measurement sample is located.
- the setting method of (3) is based on the color for the skeleton as in the setting method of (2), and the sample to be measured is not good to be separated from it, but the color to be dyed is the basic (standard) color. It is a way to see the relationship between the basic color and the actual measured value, or to match the basic color. For example, if a spider's web is used as a metaphor, in any of the setting methods (1), (2) and (3), the part where the threads of the spider's web are connected to a cross is used as the reference color.
- the sample color in the setting methods of (1) and (2), the sample color is It can be located anywhere, on the yarn, or anywhere except the yarn.
- the sample color is a color for viewing or adjusting other positional relationships based on the spider web. However, the position of the sample color should be located in the spider web.
- the setting method (3) implies setting of the reference color for the purpose of observing the positional relationship of the sample colors or for the purpose of adjusting the position using the sample colors.
- each micro color filter MF1 is the spectral spectrum that matches the spectral spectrum of the substance in the measurement sample. Is set to a color with a torque.
- each micro color filter MF1 is manufactured based on only the color value.
- the substance in the measurement sample is quantified based on a reference sample under a microscope (for example, a sample that has been dyed at a certain concentration under a certain condition), adjust to the spectral spectrum of the sample.
- a microscope for example, a sample that has been dyed at a certain concentration under a certain condition
- a color capable of emitting invisible light may be set as the color of each micro color filter MF1, It may be included in the used micro color filter MF1.
- invisible light refers to light having a wavelength in the invisible region (for example, light having a wavelength in the range of 200 to 380 nm or light having a wavelength in the range of 780 nm to 20 ⁇ ).
- colors are defined based on visible light, but in the invisible region, it is possible to express in a similar way, that is, by applying three reference functions to an arbitrary spectral distribution. For this reason, standards in the invisible region are also required and are included.
- Each micro color filter MF for color reference used for the slide glass (10, 10A, 10B) with each color reference micro filter shown in Fig. 5, Fig. 13 ⁇ and Fig. 16 ⁇ . 1 is to use a color resist ink material, which is used for color imaging devices and liquid crystal printing, as a basic color material, and to form this on a glass plate 1 using, for example, semiconductor printing technology (photomask technology). Can be. Products manufactured according to this manufacturing method have little change over time and are effective for mass production.
- a filter obtained by thinly slicing a color glass filter formed of a color developing material that forms a color in an ion or colloidal state may be used as each micro color filter MF1.
- a commercially available bandpass finoleta manufactured by Optima, trade name: "DTB # 450"
- DTB # 740 is prepared by slicing 100 to 400 ⁇ m and laminating them.
- Optima “DTB # 615” or longer wavelength bandpass filter HOYA “0-54”, “0-58”, “R-60”, “ R_62 '', ⁇ R-64 '' A cut-cut filter may be used, but if they are used, the color is too dark and does not fit into the standard color system of a camera, so it is difficult to adjust the slice thickness alone, reducing the content of coloring material, etc. Needs to be adjusted.
- a plurality of colors (at least two or more colors) be seen in the field of view at the same time as in the slide glass with the color reference microfilter according to the above-described first to third embodiments. It is more convenient to measure multiple colors (at least two or more colors) at the same time from the measurement work and data processing.
- each micro color filter MF1 As a coloring material of each micro color filter MF1, in addition to the above, a glass-specific pigment for glass printing may be used. This is obtained by mixing a metal pigment with a fine glass frit and firing at high heat (700-10000 degrees). Commercially available products include, for example, glass colorants manufactured by Okuno Pharmaceutical Co., Ltd. [0149] As another manufacturing method of each micro color filter MF1, there is a method of printing ink for a color printer on the glass plate 1 using a color printer. However, these are printed with fine dots, and when viewed under a microscope, those dots are perceived by the observer. The color obtained by a color printer is obtained by spatially integrating the dots in a certain area, and observing each dot individually will not produce the desired color.
- each micro color filter MF1 a method using an interference filter as each micro color filter MF1 is also exemplified.
- a photonic crystal may be used as a material of each micro color filter MF1. This is effective when the micro color filter MF1 is used as a color reference in the visible region and the invisible region.
- each micro color filter MF 1 is formed by using the above-described semiconductor printing technique. This will be explained.
- each micro color filter MF 1 is formed in a predetermined area on the glass plate 1 by using a photolithography technique employed for manufacturing a liquid crystal or a semiconductor element.
- each micro color filter MF1 can be manufactured through the following steps.
- the glass plate 1 to be used may be a member having a standardized size (for example, 76 mm X 26 mm) and a shape (cut) as a slide glass, or a size other than that (for example, A member having a size of 880 mm ⁇ 680 mm or 15 Omm ⁇ 15 Omm) and a shape may be used, and may be cut to a required size and shape in a later step. For example, a plurality of glass plates 1 (on which the micro color filter MF 1 has been printed) may be cut into a required size and shape.
- the glass material as a constituent material of the glass plate 1 is a target such as a slide glass. Quartz glass, low-expansion glass (aluminum borosilicate glass, borosilicate glass), soda lime glass, etc., which are used as standard, can be used, but it is better to use glass with less fluorescence.
- a substrate for liquid crystal for example, manufactured by Koning Co., Ltd., trade name:
- Color # 7059 is compatible with the color resist described below, and may be used.
- the glass plate 1 is washed first. This cleaning is usually the Shawa one wash, brush cleaning, in addition to the UV Bruno 0 3 is carried out using an ultrasonic cleaning device or the like.
- the color resist applied to the glass plate 1 to form each micro color filter MF1 is a commercially available red (R), green (G), and blue (B). Things can be used.
- R red
- G green
- B blue
- M-G302S and M-B302 manufactured by Hitachi Chemical Co., Ltd., brand name: "PD-300 series”, manufactured by Aitez Corporation, brand name: “Type 2” and “Type 3,” Dainippon Ink and Chemicals Color paste for color filters manufactured by Kogyo Co., Ltd. (trade name: “Exedic”).
- the color register for the intermediate color system between each of the three colors eg, yellow, cyan, purple (Magenta)
- a color resist is produced by adding a pigment that produces a special characteristic color.
- the prepared color resist is applied to the glass plate 1.
- a spin coater is usually used.
- slit spin coater, lono recorder, wire bar coater, die Coating may be carried out by a coater or a recently introduced CAVEX coater (Micro Giken).
- CAVEX coater Mocro Giken
- the amount of the coating solution that is discarded may be increased in some cases.
- the coating using the CAVEX coater is particularly preferable because the use ratio of the color resist is high.
- a pre-bake treatment is performed to evaporate the solvent from the applied liquid.
- a temperature of about 60 to 100 ° C and a time of about 1 to 3 hours are appropriate.
- mask exposure is performed using an exposure apparatus (such as a stepper or a mask aligner).
- an exposure apparatus such as a stepper or a mask aligner.
- a mask for example, a mask having a shape corresponding to the arrangement of each component shown in FIGS. 5, 13A, and 16A
- exposure apparatus such as a stepper or a mask aligner.
- a large-area color palette section 2 is provided so that not only color values but also spectral transmission characteristics can be accurately recorded.
- the large area color pallet unit 2 is provided with a micro color filter MF 1 provided in advance, so that the manufacturer of the micro color filter MF 1 can prepare a sample of the measurement sample in advance so that the temporal change of the spectral transmittance can be traced later if necessary. This is useful when you need to keep the spectral distribution. Also, since the spectral transmission characteristics can be easily measured, the user can easily determine the degree of fading of the micro color filter MF1 used.
- a fusion image (Fusion Image) of the image data of the measurement sample and the previously captured reference color filter image is created and displayed on the display.
- the fused image is used for recording information and / or calculating data. For example, when the user needs to perform image processing including color conversion such as saturation enhancement, hue shift, brightness adjustment, or y correction, the fused image is used.
- the measurement sample may be covered by the micro color filter MF 1 or a part of the color pallet for microscopic observation including them. Also, without using the micro color filter MF 1 which is not covered with the measurement sample or the color pallet for microscopy including the micro color filter MF 1, the photographing of this portion can be performed without any trouble.
- the absorption of the adhesive used for preparing the preparation (for example, trade name: "Canadian balsam") and the dye used for staining the sample were diluted and diluted.
- the absorption contained in the adhesive and the absorption of the cover glass add to the transmission characteristics of the micro color filter MF1, so that the value of the captured image is the sum of all these absorptions.
- the transmission characteristics of the micro color filter MF 1 are known, it is possible to calculate the absorption other than the micro color filter MF 1 from the image itself. Therefore, for example, a measurement for strictly evaluating the absorbed amount of the adhesive can be additionally performed.
- the slide glass 10B with a micro filter for color reference of Fig. 16A is a slide glass intended for use in automatic shooting, and its use method has been described in detail above. Omitted.
- the slide glass (slide glass with a color reference micro color filter) according to the present invention including the slide glasses 10, 10 A, and 10 B is a color characteristic (spectral characteristic) of the color reference micro filter.
- the colors of the images taken by different cameras and the images output by different image output means output images on a display, output images on a recording medium such as paper using a printer). Evaluation and color correction can be performed reliably.
- the image of the original measurement sample is subjected to multi-stage image processing as shown in Fig. 25. Even if this is the case, the photographing of the measurement sample and the measurement of the color reference micro-filter can be performed for each process, so that the color characteristics (spectroscopic characteristics) of the image of the original measurement sample can be processed. It is easy to understand how the change has occurred. At this time, if the light source characteristics, lens system characteristics, sensitivity characteristics, etc. at the time of shooting are also recorded for each process, a more reliable history can be obtained.
- G1 is the spectral characteristic of the reference sample having the reference color.
- a graph showing an example of (light transmittance) (for example, the graphs shown in FIGS. 7 to 12), G2 is a graph showing spectral characteristics (light transmittance) of a measurement sample, and G3 is a spectrum of a light source.
- a graph showing an example of the characteristics G 4 is a graph showing an example of the spectral characteristics (light transmittance) of the lens system, G 5 is a graph showing an example of the sensitivity characteristics (raw data) of the camera, and G 6 is a sensitivity characteristic of the camera.
- 7 is a graph (for example, s RGB system) showing an example of (correction data).
- M is It is a conversion matrix for converting G5 to G6.
- [0 1 7 7]. 7 is an example of the relationship between the measurement sample (symbol “ ⁇ ” in the figure) obtained from the color conversion performed using 01 to G6 and the reference color (symbol “ ⁇ ” in the figure). It is. G7 converts the display data of the s RGB color coordinate system to the display data of the XYZ color system for the measurement sample and the reference color, and further converts it to the display data of the a * color system. An example is shown in which the relationship between the a * value and the b * value obtained in this case is expressed.
- the slide glass with a color reference micro color filter according to the present invention when the slide glass with a color reference micro color filter according to the present invention is not used, for example, the spectral characteristics of a light source, the spectral distribution characteristics of a color camera, and the like are recorded in each processing step. Even if you leave it as it is, the slight change in the final color software on the personal computer will change the color, so the history will be almost useless, in short, It simply becomes a picture taken souvenirically and loses its meaning as a measured value. Therefore, it is considered that analytical evaluation cannot be performed later. [0179] As described above, according to the slide glass with the color reference micro color filter according to the present invention, how the color reference micro color filter is recorded, Since it is possible to know how colors are changed and expressed, for example, the following usage is effective.
- a usage method in which a pallet image of a color reference micro color filter photographed under the same conditions as a measurement sample is reduced and attached to an edge in the measurement sample image Is mentioned.
- the color-based mini-image (index image) under the same conditions is handled at the same time as the force measurement sample image. Is added. Therefore, the degree of change from the original color (color characteristics) can be checked, which proves to be extremely convenient (for example, see Fig. 26).
- G8 is a graph showing an example in which the reference value data before correction is expressed as a relationship between a * value and b * value in the L * a * b * color system.
- FIG. 9 is a graph showing an example in which the corrected reference value data is expressed as a relationship between the direct and b * values in the L * a * b * color system. Further, in G8 and G9 in FIG. 26, the symbol “ ⁇ ” indicates the data of the measurement sample, and the symbol “ ⁇ ” indicates the data of the reference color.
- the image of the measurement sample is recorded in the same manner as the image of the color reference micro color filter, the image is captured in any state when the image is later checked (measurement) You can see if it was done. Also, when color correction is performed on the collected image, if the correction is performed at the same time as the color reference image degree of the micro filter, the history of color correction remains, and how the color correction was added later It can be analyzed whether color correction has been applied. On the other hand, since the original color correction also uses the color information of the reference image portion, this color reference is essential.
- a method of using as a color matching reference can be cited.
- a system that collects an image under a microscope, including a microscope expresses the image on a display, etc., or prints the image on a printer, and records the image information, a micro color filter for color reference is photographed. Then, it can be displayed on a display, and its color can be evaluated, or printed by a printer or the like, and the color can be evaluated.
- the evaluation can be performed while viewing the actual micro color filter under visual observation.
- a color reference micro color filter that matches the photographing color with the color displayed on the display and the color expressed by the printer can be created. Color matching standards can be set.
- RGB value for simplicity, this RGB value is assumed to be R sRGB G sRGB B sRGB in the s RGB color system
- Infinite spectral distribution Ie, the presence of a metamer
- the dye of the reference measurement sample (micro-color filter for color reference) or the spectral spectrum of the dye or dye used to dye the measurement sample is known, and the unknown measurement sample is dyed.
- the dyes and dyes used for the measurement are already known, that is, if the previously prepared reference sample (color reference micro color filter) covers the dye (dye) of the unknown sample, From the measured values (for example, R, G, and B) of the photographed camera, spectral estimation of the unknown measurement sample becomes possible. Of course, quantification is also possible.
- the slide glass (slide glass with a color reference micro color filter) according to the present invention including the slide glass with a color reference micro color filter according to each of the above-described embodiments, as shown in FIG. Since the code 6 and the memo column 7 also include the application of a magnetic material or the embedding of an IC chip, various information can be recorded as a single unit.
- voice information can also be digitally recorded for memos at the time of microscopic diagnosis.
- Image information can also be digitized and recorded. Therefore, in the case of a measurement sample whose chemical state or physical state changes over time, there is-at regular intervals, the slide glass with the measurement sample is automatically removed from the storage location and automatically Measurement, and the temporal change data can be photographed and written in the memo column.
- the slide glass color reference micro
- the following effects can be obtained with a color glass slide with a color filter) by using a color reference micro color filter under a microscope.
- the use of the slide glass (slide glass with a color reference microfilter) according to the present invention is not limited to the image capturing system.
- the slide glass according to the present invention is photographed with a spectroscopic imaging sensor (for example, a spectroscopic imaging sensor described in Japanese Patent No. 2713838). You can use it as a color standard when you do.
- a spectroscopic imaging sensor for example, a spectroscopic imaging sensor described in Japanese Patent No. 2713838. You can use it as a color standard when you do.
- various information can be obtained from the acquired image data. For example, if this is used, the state of the spectrum for each part can be known, so if a basic data spectrum is prepared, it becomes possible to identify the sample containing the prepared basic data.
- the amount can be fixed. In this case, it is better to collect basic data in the same system to obtain more accurate results. Therefore, a measurement sample that becomes a reference for quantification of dyeing with a known dye concentration and dyeing bath time is required.
- the sample to be actually measured may be used as a reference, or a phantom imitating the sample may be dyed.
- two or more color reference micro color filters are arranged on the upper surface of the glass plate.
- these color reference micro color filters may be arranged on a cover glass that is in contact with the glass plate with the measurement sample interposed therebetween.
- the microscope is designed to focus on the surface of the cover glass on the measurement sample side, not on the upper surface (upper surface of the glass plate) of a normal slide glass. Also, the microscope field of view does not exceed the size of the cover glass.
- FIG. 27A is a diagram showing a basic configuration of the cover glass according to the first embodiment of the present invention
- FIG. 27B is a diagram showing a basic structure of the cover glass according to the second embodiment of the present invention. It is a figure showing a basic composition.
- the cover glass 2OA according to the first embodiment shown in FIG. 27A has a square shape, and a plurality of color reference micro color filters are provided on the surface on the measurement sample side. MF is printed.
- the cover glass 2OA according to the first embodiment is printed with an identification mark 200 for distinguishing the upper surface from the lower surface.
- the cover glass 2 OB according to the second embodiment shown in FIG. 27B is rectangular, A plurality of color reference micro color filters are arranged asymmetrically with respect to the center of gravity of the cover glass 20B. Even with such an arrangement of the color reference micro color filters, it is possible to distinguish the upper surface and the back surface of the external force bar glass 20B.
- the surfaces of the cover glasses 2OA and 2OB according to the first and second embodiments on the side of the sample to be measured had a plurality of surfaces corresponding to the respective magnifications of the objective lens mounted on the microscope.
- a color reference micro color filter MF may be arranged, and a plurality of color reference micro color filters MF may be studded all over the cover glass 20A, 20B.
- the color reference micro color filters MF are studded on the entire cover glass 20A, 20B, even if some of the color reference micro color filters MF are hidden in the measurement sample, the remaining Use of the color reference micro-color filter MF enables clear images to be captured.
- a pathological diagnosis system according to the present invention will be described.
- a slide glass silica glass according to the present invention
- a cover glass covering glass according to the present invention
- FIG. 28 is a diagram showing a schematic system configuration when performing remote pathological diagnosis as the pathological diagnostic system according to the present invention.
- This remote pathological diagnosis system is configured to perform pathological diagnosis from the observation terminal 300 constituting the image acquisition system via a wired or wireless communication line 400 such as a LAN, the Internet, a satellite line, and a telephone line.
- the configuration is such that the image data is transferred to the diagnosis-side terminal 500 that performs the processing.
- the observation-side terminal 300 in the remote pathological diagnosis system shown in Fig. 28 includes a microscope 310, a control device 320, a computer 330, an image converter 340, and a monitor 3 50 is provided.
- the microscope 310 includes an X_Y stage 311 on which the slide glass 10 and the cover glass 20 are installed as described above, and a revolver on which an objective lens 313 having a different magnification is attached.
- the control unit 3 2 0 controls the drive of the X- ⁇ stage 3 1 1 And control of image data capture.
- pathological diagnosis software is executed, and the image data (including the image of the color reference micro color filter taken for the color tone control) transmitted from the control device 320 is used.
- the image data is transmitted to the image converter 340, and transmitted from the IZ0 331 to the diagnosis terminal 550 via the communication line 400.
- the image converter 340 controls the color to be displayed on the monitor 350 correctly based on the color information about the color, brightness, etc. of the image data sent from the computer 330. Has been adjusted.
- the diagnosis terminal 500 in the remote pathological diagnosis system of FIG. 28 includes a computer 5100, an image converter 340, and a monitor 350.
- the pathological diagnosis software is also running on the computer 510, and the image data (the image of the color reference microcolor filter transferred from the observation terminal 300 via the communication line 400) ) Is received by I 0 5 1 1 and output to the image converter 5 20.
- the image converter 520 converts the transferred information about the color and brightness of the color reference micro color filter into color information that is set in advance and correctly displayed on a monitor.
- the pathological diagnosis system uses the slide glass or the cover glass on which the color reference micro color filter is arranged, and therefore, the diagnosis side terminal 5 apart from the observation side terminal 300. Even at 0, it becomes possible to obtain reference color and contrast information. As a result, the diagnosis terminal 500 in the pathological diagnosis system can also optimally adjust the color and contrast of the displayed image.
- the slide glasses 10A and 1OB according to the second and third embodiments described above are used, the reference color and contrast information are held together with the image of the measurement sample. Therefore, even if the color and contrast information of the captured image changes due to changes in the illumination system of the microscope, the camera, etc., the color information used as the reference is used not only for the diagnosis terminal 500 but also for the observation side. Even in the terminal 300, it is possible to express a correctly corrected image.
- FIG. 29 is a flowchart for describing pre-processing up to the start of remote pathological diagnosis in the pathological diagnostic system shown in FIG.
- any one of a slide glass (slide glass according to the present invention) on which a color reference color microfilter is arranged and a cover glass (cover glass according to the present invention) are provided. Is prepared (composed of a glass plate 10 and a cover glass 20) (step ST1).
- step ST2 When the prepared sample is set on the XY stage 311 of the microscope 310 (step ST2), the camera 3 is passed through the objective lens 313 having a predetermined magnification. 1 and 2 photograph the sample color reference micro color filter (step ST 3). Note that the photographing operation in the observation side terminal 300 is performed by the computer 330 through the control device 320.
- the image data of the color reference micro color filter collected as described above is displayed on the monitor 350 via the image converter 340 by the pathological diagnosis software, and ⁇ ⁇ ⁇ Sent from 3 1 1 to communication line 4 0 0 Step ST 4).
- the pathological diagnosis software executed in the computer 5 10 Is output to the image converter 520.
- the image converter 520 the color and contrast of the input image data are adjusted so as to be correctly displayed on the monitor 530 (step ST5), and the color-adjusted image is displayed on the monitor 530. (Step ST 6).
- the diagnostic terminal 500 instructs the observation terminal 300 to control the image tension and the microscope required for the diagnosis, and repeats data transmission and reception between these terminals 300 and 500.
- a diagnosis is made (step ST 7).
- the imaging device applied to the image acquisition system is not limited to a camera, and may be any component that captures a sample, such as a digital slide (virtual slide), a scanner, or an imaging unit in a virtual microscope.
- a digital slide virtual slide
- a scanner or an imaging unit in a virtual microscope.
- color information that can be used as a reference for color evaluation and color correction of an image of a measurement sample captured using a microscope can be obtained.
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- Microscoopes, Condenser (AREA)
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Abstract
Description
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Priority Applications (6)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US10/508,126 US20050142654A1 (en) | 2002-10-18 | 2003-10-17 | Slide glass, cover glass and pathologic diagnosis system |
DE60321159T DE60321159D1 (de) | 2002-10-18 | 2003-10-17 | Schiebeglas, abdeckglas und pathologisches diagnosesystem |
JP2004551190A JPWO2004044639A1 (ja) | 2002-10-18 | 2003-10-17 | スライドガラス、カバーガラス及び病理診断システム |
AU2003301924A AU2003301924A1 (en) | 2002-10-18 | 2003-10-17 | Slide glass, cover glass, and pathologic diagnosis system |
EP03756685A EP1500963B1 (en) | 2002-10-18 | 2003-10-17 | Slide glass, cover glass, and pathologic diagnosis system |
NO20051883A NO20051883L (no) | 2002-10-18 | 2005-04-18 | Objektglass, dekkglass og patologisk diagnosesystem |
Applications Claiming Priority (2)
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JP2002-304742 | 2002-10-18 | ||
JP2002304742 | 2002-10-18 |
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WO2004044639A1 true WO2004044639A1 (ja) | 2004-05-27 |
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ID=32310332
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PCT/JP2003/013325 WO2004044639A1 (ja) | 2002-10-18 | 2003-10-17 | スライドガラス、カバーガラス及び病理診断システム |
Country Status (7)
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US (1) | US20050142654A1 (ja) |
EP (1) | EP1500963B1 (ja) |
JP (1) | JPWO2004044639A1 (ja) |
AU (1) | AU2003301924A1 (ja) |
DE (1) | DE60321159D1 (ja) |
NO (1) | NO20051883L (ja) |
WO (1) | WO2004044639A1 (ja) |
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Also Published As
Publication number | Publication date |
---|---|
EP1500963B1 (en) | 2008-05-21 |
AU2003301924A1 (en) | 2004-06-03 |
US20050142654A1 (en) | 2005-06-30 |
NO20051883D0 (no) | 2005-04-18 |
EP1500963A4 (en) | 2006-04-26 |
JPWO2004044639A1 (ja) | 2006-03-16 |
EP1500963A1 (en) | 2005-01-26 |
DE60321159D1 (de) | 2008-07-03 |
NO20051883L (no) | 2005-04-18 |
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