JP2013254057A - Microscope imaging device and microscope imaging system - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a correct exposure in a short time even when changing an imaging element.SOLUTION: A microscope imaging system 1 is provided that comprises: optical element switching means 11 that switchingly disposes any optical element of a plurality of optical elements 12 having a characteristic transmitting fluorescence of a mutually different wavelength generated from a specimen 16 in an observation light path of the specimen; a color imaging element 101 that captures an observation image of the specimen via the optical element disposed in the observation light path by the optical element switching means and acquires a color image; a monochrome imaging element 102 that captures the observation image via the optical element used in acquiring the color image and acquires a monochrome image; pseudo color determination means 109 that extracts color information from the color image and determines a pseudo color corresponding to the optical element used in acquiring the color image on the basis of the color information; and pseudo color setting means 110 that sets the pseudo color determined by the pseudo color determination means to the monochrome image.

Description

  The present invention relates to a microscope imaging apparatus and a microscope imaging system.

For microscopes that capture observation images of samples by irradiating excitation light in a specific wavelength range to multiple stained samples that are placed on the microscope stage and stained with multiple fluorescent dyes, and detecting fluorescence emitted from the sample. An imaging device is known. When taking an observation image of a sample in such an imaging device for a microscope, one optimal fluorescent cube corresponding to the type of the sample or fluorescent dye is switched from a plurality of fluorescent cubes (optical elements) mounted on the turret. In some cases, an observation is performed by placing a monochromatic image of a specimen in a light path and then displaying the monochromatic image colored with a pseudo color corresponding to a fluorescent cube.
As an example of such an imaging apparatus for a microscope, Patent Document 1 previously stores a model number and a pseudo color of a plurality of fluorescent cubes in a predetermined memory in association with each other. A microscope image processing device that identifies the model number of the fluorescent cube based on the captured color observation image, reads the pseudo color corresponding to the fluorescent cube from the memory, and automatically sets the read pseudo color to a monochrome image is proposed Has been.

JP 2008-158001 A

  However, in the microscope image processing apparatus described in Patent Document 1, only the fluorescent cube stored in the memory is used to read the pseudo color corresponding to the fluorescent cube from the memory and set the pseudo color to a monochrome image. But can not use other fluorescent cubes. In recent years, fluorescent cubes having various performances have been provided, and it is desired to use these fluorescent cubes freely.

  The present invention has been made in view of the circumstances described above, and is a microscope capable of applying a wide variety of optical elements by simply determining a pseudo color corresponding to the optical element and improving convenience. It aims at providing the imaging device for microscopes, and the imaging system for microscopes.

In order to achieve the above object, the present invention provides the following means.
The present invention provides a color imaging device that acquires a color image by capturing an observation image of a sample via an optical element disposed on an observation optical path, and an optical element that is used when the color image is acquired. A monochrome imaging element that captures the observation image to obtain a monochrome image; and a pseudo color determination unit that extracts color information from the color image and determines a pseudo color corresponding to the optical element based on the color information; There is provided an imaging apparatus for a microscope comprising: pseudo color setting means for setting a pseudo color determined by the pseudo color determination means to the monochrome image.

According to the present invention, on the basis of the color information of the color image acquired by the color image sensor, a pseudo color determined in a pseudo manner corresponding to the optical element used when the color image is acquired is determined. The pseudo color is set to a monochrome image acquired by a monochrome image sensor.
That is, since the pseudo color is determined based on the color information of the color image, a wide variety of optical elements can be applied, and the convenience of the imaging apparatus for a microscope can be improved.

  In the above invention, the pseudo color determining means extracts color information from the color image, determines a color pattern corresponding to the color information, and determines a pseudo color corresponding to the optical element based on the color pattern. It is preferable.

In the above invention, it is preferable that the color pattern is a ratio of an R component, a G component, and a B component of a predetermined pixel included in the color image.
In this way, for example, a color based on the color component having the highest luminance among the color components can be determined as a pseudo color.

  In the above invention, a plurality of pseudo color patterns that are ratios of a predetermined R component, G component, and B component are stored, and a plurality of pseudo colors that are determined corresponding to the plurality of pseudo color patterns are stored in advance. A plurality of pseudo colors stored in the memory in advance, wherein the pseudo color determining means determines a color pattern based on a ratio of R component, G component and B component of a predetermined pixel included in the color image. It is preferable to determine the pseudo color based on the pseudo color pattern that matches the determined color pattern.

  In this way, by selecting one corresponding to the color pattern based on the color image from among the predetermined pseudo color patterns, the pseudo color can be immediately calculated without calculating the pseudo color each time based on the color pattern. A pseudo color can be determined.

In the above invention, it is preferable that light guiding means for switching the light from the sample to be guided to either the color image sensor or the monochrome image sensor is provided.
By doing in this way, the said color image and the said monochrome image can be acquired easily.

  The present invention is an optical element switching means for switching and arranging any one of the optical elements on the observation optical path of the sample among a plurality of optical elements having a characteristic of transmitting fluorescence in different wavelength ranges generated from the sample, A color imaging element that captures an observation image of the sample via an optical element disposed on the observation optical path by the optical element switching unit to acquire a color image, and an optical element that is used when the color image is acquired A monochrome imaging element that captures the observation image via a monochrome image to obtain a monochrome image; and a pseudo color determination that extracts color information from the color image and determines a pseudo color corresponding to the optical element based on the color information And a pseudo color setting unit that sets the pseudo color determined by the pseudo color determination unit to the monochrome image. That.

According to the present invention, the optical elements having different characteristics are appropriately switched by the optical element switching means and arranged on the observation optical path. Therefore, even when the same sample is imaged, the image is captured by the color imaging element. Each color component (for example, R component, G component, and B component) of the image to be displayed has different characteristics depending on the arranged optical elements. Therefore, based on the color information of the color image acquired by the color image sensor, a pseudo color determined in a pseudo manner corresponding to the optical element used when the color image is acquired is determined, and the pseudo color is determined as monochrome. The monochrome image acquired by the image sensor is set.
That is, since the pseudo color is determined based on the color information of the color image, a wide variety of optical elements can be applied, and the convenience of the microscope imaging system can be improved.

In the above invention, the pseudo color determining means extracts color information from the color image, determines a color pattern corresponding to the color information, and determines a pseudo color corresponding to the optical element based on the color pattern. It is preferable.
In the above invention, it is preferable that the color pattern is a ratio of an R component, a G component, and a B component of a predetermined pixel included in the color image.
In this way, for example, a color based on the color component having the highest luminance among the color components can be determined as a pseudo color.

  In the above invention, a plurality of pseudo color patterns that are ratios of a predetermined R component, G component, and B component are stored, and a plurality of pseudo colors that are determined corresponding to the plurality of pseudo color patterns are stored in advance. A plurality of pseudo colors stored in the memory in advance, wherein the pseudo color determining means determines a color pattern based on a ratio of R component, G component and B component of a predetermined pixel included in the color image. It is preferable to determine the pseudo color based on the pseudo color pattern that matches the determined color pattern.

In this way, by selecting one corresponding to the color pattern based on the color image from among the predetermined pseudo color patterns, the pseudo color can be immediately calculated without calculating the pseudo color each time based on the color pattern. A pseudo color can be determined.
In the above invention, it is preferable that light guiding means for switching the light from the sample to be guided to either the color image sensor or the monochrome image sensor is provided.
By doing in this way, the said color image and the said monochrome image can be acquired easily.

  According to the present invention, a variety of optical elements can be applied, and the convenience can be improved.

1 is an overall configuration diagram showing an imaging system for a microscope according to a first embodiment of the present invention. It is a block diagram which shows an example of a fluorescence cube (optical element). It is a figure which shows the spectral characteristic of the blue component of the fluorescence cube 12 which has the band pass filter 20B, the dichroic mirror 21B, and the absorption filter 22B. It is a figure which shows the spectral characteristic of the green component of the fluorescence cube 12 which has the band pass filter 20G, the dichroic mirror 21G, and the absorption filter 22G. It is a figure which shows the spectral characteristic of the red component of the fluorescent cube 12 with the band-pass filter 20R, the dichroic mirror 21R, and the absorption filter 22R. It is a flowchart about the determination and setting of a pseudo color by the imaging system for microscopes concerning the 1st Embodiment of this invention. It is a figure which shows an example of the display screen 200 of the monitor. It is a figure which shows an example of the display screen 200 of the monitor. It is a figure which shows an example of the display screen 200 of the monitor. It is a figure which shows the example of the histogram for every R, G, B color component produced | generated from the color image imaged using the fluorescence cube which has a spectral characteristic of a green component. It is a figure which shows an example of a pseudo color pattern. It is a figure which shows an example of the color pattern of a fluorescence cube. It is a figure which shows an example of the screen 200 displayed on the monitor. It is a figure which shows an example of the screen 200 displayed on the monitor. It is the figure which calculated | required R / G and B / G of each pixel of a color image, and was made into the graph. It is a whole block diagram which shows the imaging system for microscopes which concerns on the 2nd Embodiment of this invention. It is a flowchart about the determination and setting of a pseudo color by the imaging system for microscopes concerning the 2nd Embodiment of this invention.

[First Embodiment]
An imaging apparatus for a microscope according to a first embodiment of the present invention and an imaging system for a microscope including the imaging apparatus for the microscope will be described below with reference to the drawings.
As shown in FIG. 1, a microscope imaging system 1 according to the present embodiment is attached to a fluorescence microscope 2 and a fluorescence microscope 2, and a camera 3 (imaging device for microscope) that captures an observation image of a sample on the fluorescence microscope 2. ) And a calculation unit 4 that performs various processes such as setting of imaging conditions by the camera 3.

  The fluorescence microscope 2 includes an objective lens 10, a plurality of fluorescent cubes 12 (optical elements) mounted on a turret 11 (optical element switching means), an incident light source 13 such as a mercury lamp, and a lens barrel 14. An observation image of the placed specimen 16 (sample) can be visually observed. The camera 3 of the imaging apparatus 1 is disposed on the observation optical path a1 of the fluorescence microscope 2 at a position where an observation image from the fluorescence microscope 2 is projected.

  A plurality of fluorescent cubes 12 are mounted on the turret 11. The turret 11 can be driven by a driving means (not shown) such as a stepping motor, and is controlled by the PC 5 described later, and the fluorescent cube 12 is arranged on the observation optical path a1 according to the combination with the fluorescent dye of the specimen 16. Can be switched. In the present embodiment, the turret 11 will be described as having, for example, three types of fluorescent cubes 12 shown in FIG.

  The fluorescent cube 12 is an optical element having a characteristic of transmitting the fluorescence generated from the specimen 16, and includes a band pass filter 20, a dichroic mirror 21, and an absorption filter 22, as shown in FIG. Each fluorescent cube 12 transmits fluorescence having different wavelength characteristics, and is used in combination with an appropriate one depending on the fluorescent dye emitted from the specimen 16.

As specific examples, the spectral characteristics of three types of fluorescent cubes are shown in FIGS. That is, FIG. 3 is a diagram showing the spectral characteristics of the blue component of the fluorescent cube 12 having the bandpass filter 20B, the dichroic mirror 21B, and the absorption filter 22B, and FIG. 4 shows the bandpass filter 20G, the dichroic mirror 21G, and FIG. 5 is a diagram illustrating the spectral characteristics of the green component of the fluorescent cube 12 having the absorption filter 22G, and FIG. 5 illustrates the spectral characteristics of the red component of the fluorescent cube 12 having the bandpass filter 20R, the dichroic mirror 21R, and the absorption filter 22R. FIG.
Therefore, an image acquired by the microscope imaging apparatus 1 to be described later has different characteristics in color components depending on the fluorescent cube 12 arranged on the observation optical path a1 of the fluorescent microscope 2.

  The incident light source 13 emits illumination light of a predetermined wavelength, and the illumination light emitted from the incident light source 13 transmits only a specific wavelength component through the bandpass filter 20 of the fluorescent cube 12 and is reflected by the dichroic mirror 21 to be reflected by the objective lens 10. The sample 16 is irradiated via

  The specimen 16 is excited by the irradiated light and emits fluorescence. This fluorescence passes through the dichroic mirror 21 through the objective lens 10, passes through the absorption filter 22, and is captured as an observation image by the camera 3.

  The calculation unit 4 includes a personal computer (hereinafter simply referred to as “PC”) 5 that functions as a calculation processing unit that performs various processes, a keyboard 6 and a mouse 7 as an input unit for a user to input to the PC 5, A monitor (for example, a liquid crystal display device) 8 that displays an image captured by the camera 3 is provided.

  The PC 5 includes a memory 111, a color image processing unit 109 (pseudo color determining unit) and a monochrome image processing unit including a CPU (central processing unit) built in the PC 5 to execute a program stored in the PC 5 in advance. 110 (pseudo color setting means). The memory 111 stores a plurality of pseudo color patterns which are ratios of a predetermined R component, G component and B component, and a plurality of pseudo colors corresponding to the pseudo color patterns (see FIG. 11).

  The color image processing unit 109 generates a color image based on a digital color image signal output via an I / F unit 108 of the camera 3 to be described later, and also generates a memory 111 based on the digital color image signal. The pseudo color is determined with reference to the pseudo color pattern stored in. That is, the color image processing unit 109 extracts color information from a digital color image signal, determines a color pattern corresponding to the optical element used when the color image signal is acquired based on the color information, and A pseudo color corresponding to the optical element is determined in correspondence with the color pattern.

  The monochrome image processing unit 110 generates a monochrome image based on the digital monochrome image signal output via the I / F unit 108 of the camera 3 and converts the pseudo color calculated by the color image processing unit 109 into a monochrome image. Set.

  That is, the PC 5 outputs various instructions and imaging conditions input from the keyboard 6 and the mouse 7 to the camera 3, selects a pseudo color of the fluorescent cube used based on the captured image, and is sent from the camera 3. An image obtained by superimposing the pseudo color for each selected fluorescent cube 12 on the monochrome image is displayed on the monitor 8.

  The microscope imaging apparatus 1 includes a prism 100, a color imaging device 101, a monochrome imaging device 102, pre-processing units 103 and 104, a timing generator (hereinafter simply referred to as “TG”) 105, an imaging device control unit 106, and a control unit 107. , An interface unit (hereinafter simply referred to as “I / F unit”) 108.

The prism 100 is an optical element that switches the observation optical path a1, and is an example of a light guide unit that switches and guides subject light to the color imaging element 101 or the monochrome imaging element 102.
For the color image sensor 101 and the monochrome image sensor 102, for example, an image sensor such as a CCD or a CMOS can be applied. The color image sensor 101 has a color filter formed on the light receiving surface thereof, and obtains a color image by capturing an observation image of the specimen 16 via any fluorescent cube 12 arranged on the observation optical path a1. . In addition, the monochrome imaging element 102 does not have a color filter formed on its light receiving surface, and captures an observation image of the specimen 16 through any fluorescent cube 12 arranged on the observation optical path a1 to obtain a monochrome image. To get.

  In this embodiment, red (R), green (G), and blue (B) primary color filters are regularly arranged for each pixel as color filters formed on the light receiving surface of the color image sensor 101. Let it be a color filter. In this case, for example, a Bayer color filter can be employed as the color filter. The color image sensor 101 and the monochrome image sensor 102 are arranged so as to image the same portion of the sample 16.

  The color pre-processing unit 103 uses an output signal of the color image sensor 101 as an image signal, amplifies the image signal, and then converts the amplified image signal into a digital color image signal. The monochrome pre-processing unit 104 uses the output signal of the monochrome image sensor 102 as an image signal, amplifies the image signal, and then converts the amplified image signal into a digital monochrome image signal.

The TG 105 is controlled by an image sensor control unit 106 to be described later, and generates pulses necessary for controlling the color image sensor 101, the monochrome image sensor 102, the color pre-processing unit 103, and the monochrome pre-processing unit 104.
The image sensor control unit 106 instructs the color image sensor 101 and the monochrome image sensor 102 to shoot in accordance with an instruction from the PC 5 instructed via the I / F unit 108, and sets the exposure time and gain set by the PC 5. Accordingly, the TG 105, the color pre-processing unit 103, and the monochrome pre-processing unit 104 are controlled.

The control unit 107 outputs the digital signals output from the color pre-processing unit 103 and the monochrome pre-processing unit 104 to the PC 5 in accordance with an instruction from the PC 5 instructed via the I / F unit 108.
The I / F unit 108 outputs the digital signals output from the color pre-processing unit 103 and the monochrome pre-processing unit 104 to the PC 5 in accordance with an instruction from the control unit 107, and outputs the instruction from the PC 5 to the control unit 107. To do.

  When the specimen 16 is a multiple-stained specimen stained with a plurality of fluorescent dyes, in the fluorescent microscope 2 described above, the optimum fluorescent cube 12 corresponding to the type of fluorescent dye is switched and placed on the observation optical path a1. The specimen 16 is observed. In particular, when performing fluorescence observation, the sample 16 may be photographed with high sensitivity or a monochrome image in consideration of the quantitativeness of the photographed image. When capturing a monochrome image, the microscope imaging system displays an image captured for each fluorescent cube 12 in color by superimposing the image on the monochrome image for observation. That is, the color image processing unit 109 determines the pseudo color based on the digital color image signal, sets the monochrome image with the determined pseudo color, that is, colors the monochrome image, and displays it on the monitor 8.

Hereinafter, according to the flowchart of FIG. 6, the determination and setting of the pseudo color by the microscope imaging system according to the present embodiment will be described.
In step S1, a program stored in advance in the PC 5 is activated, and a screen 200 (see FIG. 7 and the like) based on this program is displayed on the monitor 8.
Here, the screen 200 includes an image display unit 201, a live shooting button 202, a still image shooting button 203, a fluorescent cube switching unit 204, a color image / monochrome image acquisition switching button 208, and a reset button 210. The fluorescent cube switching unit 204 includes three types of fluorescent cube switching buttons 205, 206, and 207 set on the turret 11. The live shooting button 202, the still image shooting button 203, the fluorescent cube switching buttons 205, 206, 207, the color image / monochrome image acquisition switching button 208 and the reset button 210 can be operated with the keyboard 6 and the mouse 7. ing.

For example, when the live shooting button 202 is clicked (pressed) once with the mouse 7, the imaging apparatus 1 starts live shooting, a live image is displayed on the image display unit 201, and when clicked again, the live shooting ends. Yes.
When the still image shooting button 203 is clicked with the mouse 7, the still image shooting is performed once, and the shot image is displayed on the image display unit 201. When the fluorescent cube switching buttons 205, 206, and 207 are clicked with the mouse 7, the turret is rotated, and the clicked fluorescent cube is set on the observation optical path a1. The color image / monochrome image acquisition switching button 208 controls the prism 100 to switch the observation optical path a1, that is, to switch the subject light to the color image sensor 101 or the monochrome image sensor 102.

  When the program is started or the reset button 210 is clicked (pressed) in step S1, color image capturing processing is started for each fluorescent cube 12 in step S2. That is, in step S2, the turret 11 is rotated, all the fluorescent cubes 12 set in the turret 11 are set in order on the observation optical path a1, and the sample 16 is obtained by the color imaging device 101 using each fluorescent cube 12. An observation image is taken. Then, the color pre-processing unit 103 generates a digital color image signal from the image signal captured by the color image sensor 101.

  In the next step S3, the color image processing unit 109 generates a color image. That is, in the previous step S 2, a color image is generated in the color image processing unit 109 based on the digital color image signal generated by the color pre-processing unit 103, and the generated image is displayed on the monitor 8. 7, 8, and 9, when the three types of fluorescent cubes 12 set on the turret 7 are the fluorescent cube A, fluorescent cube B, and fluorescent cube C, color images captured using the fluorescent cubes 12 are shown. An example of the displayed screen 200 is shown.

  In the next step S4, the PC 5 generates a histogram for each R, G, B color component for each generated color image. FIG. 10 shows an example of a histogram for each of the R, G, and B color components generated from the color image captured using the fluorescent cube B. In FIG. 10, the horizontal axis represents gradation, the vertical axis represents frequency, and the histogram is generated for all pixels of one color image.

  In step S5, as shown in FIG. 10, in order not to be affected by noise or the like, the maximum gradation value excluding the upper 10% frequency of the histogram generated in step S4 is set to R, G, B colors. Calculate for each component.

  In step S6, the maximum gradation values (hereinafter referred to as “first maximum values”) calculated for each of the R, G, and B color components calculated in step S5 are compared, and the maximum value (hereinafter referred to as “first”) is compared. 2) is calculated. By doing in this way, the color pattern corresponding to the fluorescence cube used when the color image was imaged can be grasped. In the case of FIG. 10, comparing the maximum values of the R component, G component, and B component gradations, the R component gradation is 30, the G component gradation is 100, and the B component gradation is 10. Select ingredients.

In the next step S7, one of the pixels having the color component selected in step S6 is selected from the pixels having the same gradation value as that of the second maximum value. In the case of FIG. 10, since the gradation of the G component selected in step S6 is 100, one pixel having a gradation with a G component of 100 is selected.
In step S8, the ratio of the R component, G component, and B component of the pixel selected in step S7 is obtained. The ratio of the R component, G component, and B component is R / M, G / M, and B / M, where M is the maximum value of the R component, G component, and B component of the selected pixel.

  In step S9, R / M, G / M, and B / M calculated as a ratio of the R component, G component, and B component determined in step S8 are 1 or less than a predetermined constant K, less than the constant K. The color pattern corresponding to the fluorescent cube 12 is determined with the value of 0 as 0. Then, a pseudo color pattern that matches the determined color pattern is selected from the pseudo color patterns (for example, FIG. 11) stored in the memory of the PC 5, and each fluorescent cube 12 and the selected pseudo color pattern are associated with each other. Store in memory.

  In the case of FIG. 10, the G component is the maximum value, and M = 100. Here, if K = 0.5, R / M = 0.3 (<0.5), G / M = 1 (> 0.5), B / M = 0.1 (<0.5) Therefore, the color pattern of the fluorescent cube B is R = 0, G = 1, and B = 0. Therefore, a pseudo color pattern with R = 0, G = 1, and B = 0 is selected from the prepared pseudo color patterns. By doing so, it is possible to grasp the spectral characteristics of the fluorescent cube assumed in the previous step S6 as a color pattern, and to determine a pseudo color pattern for an image captured using the fluorescent cube.

  FIG. 11 shows an example of the pseudo color pattern stored in the PC 5 in advance, and FIG. 12 shows the case where the three fluorescent cubes 12 are fluorescent cubes A, B, and C. An example of the color pattern of each fluorescent cube A, B, C obtained from the ratio of As described above, since the color pattern of the fluorescent cube B is R = 0, G = 1, and B = 0, the pseudo color pattern of the fluorescent cube B is the pseudo color pattern 2 of the pseudo color patterns shown in FIG. Will be applicable.

  In step S10, first, an observation image is captured by the monochrome imaging element 102 via the fluorescent cube used in capturing the color image in step S2, and an image signal is acquired. The monochrome image processing unit 110 generates a monochrome image based on the digital monochrome image signal digitized in step S1. Then, the pseudo color corresponding to the pseudo color pattern selected in the previous step S9 is set in the monochrome image, that is, by superimposing the image colored with the pseudo color on the monochrome image, the pseudo color is applied to the monochrome image. Color.

  In step S11, a monochrome image in which a pseudo color is set is displayed on the image display unit 201. As the pseudo color, the value of the monochrome image data itself may be converted into an RGB value, or the pseudo color may be added to the image data as pseudo color information and set only when displayed on the screen 200.

  Further, on the screen 200, the superimposed display is performed by operating the fluorescent cube switching buttons 205, 206, and 207 on the screen 200 to switch the fluorescent cube 12 to the three monochrome images photographed, and for each used fluorescent cube shown in FIG. Corresponding pseudo colors can be automatically set and overlaid. FIG. 13 shows a screen 200 on which the superimposed image is displayed.

  As described above, according to the imaging system for a microscope according to the present embodiment, based on the information of each color component of the color image acquired by the color imaging element 101, the fluorescent cube used when the color image is acquired. Identify the corresponding color pattern, determine the pseudo color determined in accordance with the determined color pattern, and set this pseudo color to the monochrome image acquired by the monochrome image sensor, so it is easier to set the pseudo color it can. In other words, even with a fluorescent cube that has never been used before, the color pattern corresponding to the fluorescent cube can be determined each time, so the pseudo color corresponding to the color pattern is determined, and a wide variety of fluorescent cubes are determined. A cube can be applied, and the convenience of the imaging system for a microscope can be improved. Further, by automatically calculating the pseudo color and setting the monochrome image in the PC 5, it is possible to reduce the burden of the operator setting the pseudo color manually.

In the above-described example, as shown in FIG. 6, when the screen 200 is activated and when the reset button 210 is clicked, a color image is captured for each fluorescent cube 12 set on the turret 11, and fluorescence is captured from each captured color image. Although the pseudo color corresponding to the cube 12 has been calculated, the color image is displayed each time the fluorescent cube 12 placed on the observation light path a1 is switched by rotating the turret 11 instead of when the screen 200 is activated and when the reset button 210 is clicked. The pseudo color may be calculated from the color image.
By doing so, since the pseudo color is recalculated every time the fluorescent cube 12 is switched, a more accurate pseudo color can be set.

Alternatively, every time a monochrome image is taken, a color image may be taken, and a monochrome image may be taken after calculating a pseudo color. In this case, the fluorescence of the fluorescent cube 12 or the like being used may be taken. Even if there is no information related to the microscope, it is possible to always set an accurate pseudo color.
At this time, it is preferable to calculate the pseudo color by capturing the color image by capturing the monochrome image, for example, by retracting the prism 100 from the observation optical path a1. By operating in this order, the pseudo color can be set for the monochrome image when the operator presses the shooting button.

  The turret 11 can be configured to be switched automatically by an operation from the PC 5 or can be switched manually. In this way, the pseudo color can be set not only for the electric microscope imaging system but also for the manual microscope imaging system.

The histogram generated for the pseudo color calculation is intended for all pixels of the color image, but is not necessarily limited to this. In order to generate a histogram as shown in FIG. It is also possible to select the histogram generation area 209 and generate a histogram and calculate a pseudo color based on the histogram generation area 209.
By doing this, it is possible to select an optimal region for pseudo color calculation, reduce the number of pixels necessary for generating a histogram, and reduce the amount of calculation for pseudo color calculation. The pseudo color can be calculated faster and more accurately.

In the above example, a histogram is generated to calculate the ratio of R component, G component, and B component for pseudo color calculation. The ratio of R component, G component, and B component is shown in FIG. Thus, R / G and B / G of each pixel may be obtained and graphed, and the ratio of the R component, G component, and B component of the center of gravity may be calculated.
Alternatively, the operator may select a pixel to be used for pseudo color calculation from the color image, and calculate the pseudo color from the ratio of the R component, G component, and B component of the selected pixel.
In this way, a more accurate pseudo color can be calculated. Further, when the operator selects a pixel to be used for pseudo color calculation, more accurate pseudo color can be set.

  In the above-described example, only the epi-illumination light source 9 is provided, but a configuration including a transmission illumination light source may be employed. In this case, in step S2 in FIG. 6, when the color image sensor 101 captures an image, light emitted from the transmitted illumination light source and transmitted through the fluorescent cube 12 may be captured. By using the transmitted illumination light source, the pseudo color can be set stably and accurately regardless of the presence or absence of the specimen.

  In the above example, a pseudo color is calculated by calculating a color pattern once based on the ratio of the R component, G component, and B component calculated from the color image, and applying this to the pseudo color pattern stored in the memory in advance. However, a pseudo color can be set from the color pattern itself calculated from the color image. In this way, more accurate pseudo color setting can be performed.

[Second Embodiment]
The microscope imaging system according to the second embodiment of the present invention will be described below with reference to the drawings.
In the above-described first embodiment, the color image processing unit 109, the monochrome image processing unit 110, and the memory 111 are provided in the PC 5, but as shown in FIG. 16, the camera 3 (imaging device for microscope) includes them. It can also be set as the structure provided.

In this case, the color image processing unit 109 generates a color image based on the digital signal output from the color preprocessing unit 103, and selects a pseudo color pattern from the generated color image. The monochrome image processing unit 110 generates a monochrome image based on the digital signal output from the monochrome preprocessing unit 104, and adds the pseudo color pattern information read from the memory 111 to the monochrome image.
The memory 111 stores the color image generated by the color image processing unit 109, the histogram, and the selected pseudo color pattern.
In addition, the same code | symbol is attached | subjected to the structure same as 1st Embodiment mentioned above, and the description is abbreviate | omitted.

The pseudo color determination and setting by the microscope imaging apparatus according to the present embodiment will be described with reference to the flowchart of FIG.
When the program is started or the reset button 210 is clicked (pressed) in step S21, a color image is captured for each fluorescent cube 12 in step S22, and a color image is generated in the color image processing unit 109 in step S23. In step S24, the color image generated by the color image processing unit 109 is transferred to the memory 111 and stored therein.

  In the next step S25, the color image processing unit 109 generates a histogram for each of the R, G, and B color components for each generated color image. In step S26, the color image processing unit 109 is not affected by noise or the like. As in the example shown in FIG. 10, the maximum value of the gradation excluding the upper 10% frequency of the histogram is calculated for each of the R, G, and B color components.

  In step S27, the maximum gradation values for the R, G, and B color components calculated in step S26 are compared, and the maximum value is calculated. In the next step S28, the color image stored in the memory 111 is read, and in step S29, the color component is selected from the pixels having the color component selected in step S27 out of the color image read in the previous step S28. One pixel having one of the same gradation values as the second maximum value is selected. In step S30, the ratio of the R component, G component, and B component of the pixel selected in step S29 is obtained. The ratio of the R component, G component, and B component is R / M, G / M, and B / M, where M is the maximum value of the R component, G component, and B component of the selected pixel.

  In step S31, R / M, G / M, and B / M obtained in step S30 correspond to the fluorescent cube 12 with 1 being a value greater than or equal to the predetermined constant K and 0 being less than the constant K. Determine the color pattern to be used. Then, a pseudo color pattern that matches the determined color pattern is selected from the pseudo color patterns stored in advance in the color image processing unit 109. The selection of the pseudo color pattern is the same as the selection method in the first embodiment. The selected pseudo color pattern is transferred to the memory 111 and stored.

In step S32, the monochrome image processing unit 110 reads the pseudo color pattern stored in the memory 111, adds the corresponding pseudo color to the generated monochrome image, and outputs it to the I / F unit 108.
In step S33, a monochrome image in which a pseudo color is set is displayed on the image display unit 201.

The pseudo color setting is not performed by adding a pseudo color pattern to the monochrome image in the monochrome image processing unit 110, but by converting the pixel data value itself of the monochrome image to RGB based on the pseudo color pattern selected in step S31. It may be converted to a value.
As described above, according to the microscope imaging system according to the present embodiment, it is possible to set the pseudo color more easily with only the camera 3 without performing the process for setting the pseudo color in the PC 5.

  As described above, the embodiments of the present invention have been described with reference to the drawings. However, the microscope imaging apparatus and the microscope imaging system to which the present invention is applied have the above-described embodiments within the range in which the functions are executed. Various modifications are possible without being limited to forms and the like.

  For example, in the microscope imaging device and the microscope imaging system of each embodiment, processing can be performed via a network such as a LAN or WAN, whether it is a single device, a system composed of a plurality of devices, or an integrated device. It goes without saying that the system may be implemented.

  In addition, for example, in the microscope imaging system and the microscope imaging device according to each of the above embodiments, other optical elements that can divide the observation optical path a1 into two, such as a half mirror, may be applied instead of the prism 100. Is possible.

  In addition, for example, in the microscope imaging system according to the first embodiment, the function of each embodiment described above is realized by executing the program code read out on the memory by the computer, and the instruction of the program code Based on the above, the OS or the like running on the computer performs part or all of the actual processing, and the functions of the above-described embodiments are also realized by the processing.

Furthermore, a program code read from a portable storage medium or a program (data) provided by a program provider is provided in a function expansion board inserted into a computer or a function expansion unit connected to a computer. After that, the CPU of the function expansion board or function expansion unit performs part or all of the actual processing based on the instruction of the program code, and the functions of the above-described embodiments are also performed by the processing. Can be realized.
That is, the present invention is not limited to the above-described embodiments and the like, and can take various configurations or shapes without departing from the gist of the present invention.

  In this embodiment, the case of switching from the color mode to the monochrome mode has been described as an example. However, in the reverse case, the exposure time can be easily calculated by the same process and set in a short time. There is an advantage.

DESCRIPTION OF SYMBOLS 1 Imaging system for microscopes 2 Fluorescence microscope 3 Camera 4 Calculation part 5 PC
11 Turret (Optical element switching means)
12 Fluorescent cube (optical element)
16 specimens
DESCRIPTION OF SYMBOLS 101 Color image sensor 102 Monochrome image sensor 109 Color image processing part (pseudo-color determination means)
110 Monochrome image processing unit (pseudo color setting means)
111 memory

Claims (10)

A color imaging element that captures a color image by capturing an observation image of the sample via an optical element disposed on the observation optical path;
A monochrome imaging element that captures the observation image via the optical element used when acquiring the color image to acquire a monochrome image;
Pseudo color determination means for extracting color information from the color image and determining a pseudo color corresponding to the optical element based on the color information;
An imaging apparatus for a microscope, comprising: pseudo color setting means for setting the pseudo color determined by the pseudo color determination means to the monochrome image. The pseudo color determining means is
The imaging apparatus for a microscope according to claim 1, wherein color information is extracted from the color image, a color pattern corresponding to the color information is determined, and a pseudo color corresponding to the optical element is determined based on the color pattern.   The microscope imaging apparatus according to claim 2, wherein the color pattern is a ratio of an R component, a G component, and a B component of a predetermined pixel included in the color image. A plurality of pseudo color patterns that are ratios of a predetermined R component, G component, and B component are stored, and a memory that stores a plurality of pseudo colors determined in correspondence with the plurality of pseudo color patterns is provided.
The pseudo color determining means is
A color pattern is determined based on a ratio of R component, G component, and B component of a predetermined pixel included in the color image, and matches with the determined color pattern among a plurality of pseudo color patterns stored in advance in the memory The imaging apparatus for a microscope according to claim 1, wherein the pseudo color is determined based on the pseudo color pattern to be performed.   5. The microscope imaging apparatus according to claim 1, further comprising a light guiding unit that switches light from the sample to guide either the color imaging element or the monochrome imaging element. 6. An optical element switching unit that switches and arranges one of the optical elements on the observation optical path of the sample, among a plurality of optical elements having a characteristic of transmitting fluorescence in different wavelength ranges generated from the sample;
A color image pickup device for picking up an observation image of the sample through an optical element arranged on the observation optical path by the optical element switching means to obtain a color image;
A monochrome imaging element that captures the observation image via the optical element used when acquiring the color image to acquire a monochrome image;
Pseudo color determination means for extracting color information from the color image and determining a pseudo color corresponding to the optical element based on the color information;
An imaging system for a microscope comprising: pseudo color setting means for setting the pseudo color determined by the pseudo color determination means to the monochrome image. The pseudo color determining means is
The microscope imaging system according to claim 6, wherein color information is extracted from the color image, a color pattern corresponding to the color information is determined, and a pseudo color corresponding to the optical element is determined based on the color pattern.   The microscope imaging system according to claim 7, wherein the color pattern is a ratio of an R component, a G component, and a B component of a predetermined pixel included in the color image. A plurality of pseudo color patterns that are ratios of a predetermined R component, G component, and B component are stored, and a memory that stores a plurality of pseudo colors determined in correspondence with the plurality of pseudo color patterns is provided.
The pseudo color determining means is
A color pattern is determined based on a ratio of R component, G component, and B component of a predetermined pixel included in the color image, and matches with the determined color pattern among a plurality of pseudo color patterns stored in advance in the memory The microscope imaging system according to claim 6, wherein the pseudo color is determined based on the pseudo color pattern to be performed. 10. The microscope imaging system according to claim 6, further comprising a light guiding unit that switches light from the sample to guide either the color imaging device or the monochrome imaging device. 11.

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