JPH11202216A - Photographic apparatus - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a photographic apparatus which can select an optimum exposure time corresponding to microscopy at all the time. SOLUTION: The distribution of a luminance histogram is calculated from a digital voltage signal corresponding to the quantity of received light through an image sensor 10 and either of bright field microscopy or fluorescent microscopy is discriminated from the result of comparison between the distribution of the luminance histogram based on the bright field microscopy and a boundary distribution value set based on the distribution of the luminance histogram based on the fluorescent microscopy. From this discriminated result, the exposure time is operated by using an exposure reference value stored in a ROM 12 corresponding to the bright field microscopy or the fluorescent microscopy.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a microscope photographing apparatus for photographing a microscope observation image.

[0002]

2. Description of the Related Art Conventionally, bright field microscopy, fluorescence microscopy and the like have been known as typical methods for microscopic examination of a specimen. In order to record a specimen image observed under a microscope under such a microscopic method, a photograph of the specimen image observed under a microscope is also taken, and in this photography, a high-quality specimen photograph is obtained. For this reason, an optimal exposure time for the light amount of the sample image is determined.

As a method for determining such an exposure time, as disclosed in Japanese Patent Application Laid-Open No. 56-52730, an output signal of a light receiving element which generates an output signal corresponding to the brightness of a subject to be photographed is integrated. In some cases, the integration time is compared with a preset reference value, and the time until a match is obtained is defined as the exposure time.

[0004]

However, in general, the exposure criterion for photographing an observation image of a microscope is set to a specimen to be observed by a transmission bright-field microscopy method, for example, an HE-stained specimen. If the reference value is fixed as above, if the specimen is changed from brightfield microscopy to fluorescence microscopy and the specimen is photographed, the exposure standard will not match, and a good quality specimen will be obtained. Images cannot be taken. For this reason, the photographer needs troublesome work such as correcting the exposure standard every time the microscopic method is switched or switching the exposure standard according to the microscopic method, especially when the fluorescent microscopic method is selected, Skills are required to select the optimal exposure time for the photographer, such as performing a so-called test photography in which many specimen images are taken while changing the exposure time. There was also the problem of being done.

Therefore, as disclosed in Japanese Patent Application Laid-Open No. Hei 5-113589, a speculative method has been used based on a light receiving signal of a light receiving element, focusing on a fading phenomenon of fluorescence (a phenomenon in which the fluorescence intensity decreases with time). There has been proposed a method in which the exposure time is automatically determined and the exposure time is set based on the result.

However, recently, due to the progress of anti-fading agents and the progress of excitation light illumination systems, there has been a tendency to use an observation system that hardly causes fading of a fluorescent specimen. Based on this, there was a problem that the microscopic method could not be determined.

The present invention has been made in view of the above circumstances, and an object of the present invention is to provide a microscope photographing apparatus which can always set an optimum exposure time corresponding to a microscopic method.

[0008]

According to the first aspect of the present invention,
In a microscope photographing apparatus for photographing a sample image taken through an observation optical system of a microscope, a light receiving means for receiving light transmitted through the observation optical system, and respective exposure reference values corresponding to different microscopy methods Storage means for storing the information, a speculum method discriminating means for discriminating a speculum method using the variance of the luminance histogram according to the output of the light receiving means, and a speculum method discriminated by the speculum method discriminating means. And an exposure time calculating means for calculating the exposure time based on the exposure reference value.

According to a second aspect of the present invention, in the first aspect, the speculum discriminating means includes a boundary variance value set based on a variance of each luminance histogram obtained by a different speculum method and an output of the light receiving means. The microscopy method is determined from the result of comparison with the variance of the luminance histogram according to the above.

According to a third aspect of the present invention, in the first aspect, the speculum method discriminating means sets the boundary variance range using a boundary variance value set based on the variance of each luminance histogram obtained by different speculum methods. When the determination is made and the variance of the luminance histogram according to the output of the light receiving means is included in the boundary variance range, a warning means for issuing a warning prompting selection of the microscopic method is provided.

As a result, according to the first aspect, when an observation system that does not cause fading of a fluorescent specimen is used by determining a difference in an image due to a difference in a microscopic method by focusing on a variance of a luminance histogram. Also, the discrimination of the microscopic method can be reliably performed.

According to the second aspect of the present invention, it is possible to determine a highly accurate microscopic method by using the boundary variance value set based on the variance of each luminance histogram obtained by different microscopic methods. According to the third aspect, even when using a special microscopic method in which the dispersion is uneven, the microscopic method can be reliably selected.

[0013]

Embodiments of the present invention will be described below with reference to the drawings. (First Embodiment) FIG. 1 shows a schematic configuration of a microscope photographing apparatus to which the present invention is applied. In the figure,
Reference numeral 1 denotes a CPU, to which a shutter open / close drive circuit 2 is connected, and to which a shutter 3 is connected. The shutter opening / closing drive circuit 2 has a function of driving the opening / closing of the shutter 3.

A film winding motor drive circuit 4 is connected to the CPU 1, and a camera 5 is connected to the film winding motor drive circuit 4. The film winding motor drive circuit 4 has a function of driving a film winding motor 6 of the camera 5 to wind a photographic film.

A switch input circuit 7 and a display circuit 8 are connected to the CPU 1. The switch input circuit 7 is for inputting film characteristic information including at least an ISO value. The display circuit 8 includes, for example, an LCD display, a plasma display, or an LED, and displays at least an identification display and an exposure time according to the microscopic method.

The CPU 1 includes a timing generator 9
And the image sensor 10 is connected to the timing generator 9. In addition, this image sensor 1
0 is connected to the CPU 1 via the A / D conversion circuit 11. The timing generator 9 includes an image sensor 10
The timing for reading the output from the CPU is given.
The image sensor 10 includes, for example, a photoelectric conversion element such as a CCD, receives light transmitted through an objective lens of an observation optical system of a microscope (not shown), and outputs a current corresponding to the amount of received light. The current output from the image sensor 10 is converted into a digital voltage signal by the A / D conversion circuit 11 and is provided to the CPU 1.

The CPU 1 has a ROM 12 and a RAM 1
3 are connected. The ROM 12 stores a control program of the CPU 1 and an exposure reference value and a boundary variance Th of the bright field microscopy and the fluorescence microscopy in the microscope, respectively. Here, as shown in FIG. 2, for example, an exposure reference value K1 is set for the bright field microscopy method and an exposure reference value K2 is set for the bright field microscopy method. I have. The boundary variance Th is determined from the difference in the luminance distribution of the observed image between the bright field microscopy and the fluorescence microscopy.

FIG. 8A and FIG. 8B show typical luminance distributions of a bright-field image and a fluorescent image in a histogram.
In the bright-field image shown in FIG. 6A, the distribution is centered on the intermediate level, whereas in the fluorescent image shown in FIG.
It is mainly distributed at both ends, and the middle level tends to decrease. Such a difference between the histograms can be evaluated by the variance representing the variation. Generally, it can be said that the variance of the fluorescent image is larger than the bright-field image.

FIG. 3 shows, for example, 35 used for photography.
The difference in the dispersion between the bright field image and the fluorescent image on the mm film surface is shown. Here, the boundary variance Th is set at the boundary between the two as shown in the figure. By comparing the magnitude relationship between the variance calculated from the actual observation image and the boundary variance value Th, it is possible to determine whether the method is bright-field microscopy or fluorescence microscopy.

The CPU 1 includes an image sensor 10
The illuminance L is obtained from the digital voltage signal corresponding to the current output of the luminous intensity, and the variance of the luminance histogram is obtained. The exposure reference value K1 or K2 for the mirror method is selected, and the exposure time is further calculated. The variance of the luminance histogram is calculated as follows. Here, when the image sensor 10 has n × m pixels and the pixel is (i, j), the variance is represented by the following equation. Then, the variance of the image is calculated from the digital signal representing each pixel according to the current output of the image sensor 10 by using this equation.

[0021]

(Equation 1)

Next, the operation of the first embodiment configured as described above will be described with reference to the flowchart of FIG. First, in step 401, it is determined whether or not the shutter has been released by the switch input circuit 7. If NO here, it is determined in step 402 whether the ISO value has been input from the switch input circuit 7. If the ISO value has been input, in step 403, the ISO value is stored in the RAM 13, and the process proceeds to step 404. If the ISO value has been input, the process immediately proceeds to step 404.

In step 404, the illuminance L is calculated. In this case, when a part of the luminous flux transmitted through the objective lens of the observation optical system of the microscope (not shown) enters the image sensor 10, the image sensor 10 outputs a current corresponding to the amount of received light and outputs this current output to the timing generator. The data is output to the A / D conversion circuit 11 at the read timing of No. 9, where it is converted into a digital voltage signal and supplied to the CPU 1. Thereby, the CPU 1 obtains the illuminance L from the digital voltage signal corresponding to the output of the image sensor 10. Further, the CPU 1 determines in step 405 that the image sensor 1
From the digital voltage signal corresponding to the output of 0, a luminance histogram and its variance s are obtained.

Next, in step 406, the CPU 1
The variance σs of the luminance histogram is compared with the boundary variance value Th stored in the ROM 12 to determine the microscopic method. In this case, if the variance σs of the luminance histogram> the boundary variance Th, it is determined to be the fluorescence microscopy method, and the variance σs of the luminance histogram is determined.
If ≦ the boundary variance Th, it is determined to be the bright-field microscopy. Here, if it is determined that the method is the fluorescence microscopy method, at step 407, R
When the exposure reference value K2 is selected and read from the OM 12 and the bright field microscopy is determined, in step 408, the exposure reference value K1 is selected and read.

Next, at step 409, an identification display corresponding to the determined microscopic method is displayed on the display circuit 8. FIG. 5 (a)
FIG. 5B shows an example of identification display on the display circuit 8. In FIG. 5A, “AUT” for identifying the bright-field microscopy is shown.
O "is displayed, and in FIG. 5B," FL-AUTO "for identifying the fluorescence microscopy is displayed.

Next, at step 410, the CPU 1
The ISO value stored in the RAM 13 is read, and in step 411, the exposure time is calculated. In this case, the exposure time T
Is calculated from the following equation.

T = K / (AL) where K (K1 or K2) is an exposure reference value and A is an ISO
The value, L, is the illuminance. Then, in step 412, the calculated exposure time T is displayed on the display circuit 8. In this case, as shown in FIGS. 5A and 5B, the exposure time T = 0.695 seconds is displayed in both the bright field microscopy and the fluorescence microscopy.

Next, the process returns to step 401, and thereafter, the above-described operation is repeated while performing the release determination and the ISO value input determination. Thereafter, in step 401, when it is determined that the release is performed by the switch input circuit 7, in step 413, the shutter 3 is opened by the shutter opening / closing drive circuit 2, and in step 414, it is determined that the exposure time has elapsed. Then, the shutter 3 is closed by the shutter opening / closing drive circuit 2, and then step 416 is performed.
Thus, the film winding motor drive circuit 4 drives the film winding motor 6 of the camera 5 to wind the photographic film.

Therefore, according to this configuration, the variance s of the luminance histogram is obtained from the digital voltage signal corresponding to the amount of light received by the image sensor 10, and the variance of the luminance histogram by the bright-field microscopy and the luminance histogram by the fluorescence microscopy are obtained. From a comparison result with the boundary variance value Th set based on the variance of the light field microscope or the fluorescence microscopy method. Based on the result of the determination, the RO is determined according to the bright field microscopy method or the fluorescence microscopy method.
Since the exposure time is calculated using the exposure reference values K1 and K2 stored in M12, the difference between the typical image of the bright-field microscopy and the fluorescence microscopy is focused on the variance of the luminance histogram. Even when an observation system that does not cause fading of the fluorescent specimen is used, the distinction between bright field microscopy and fluorescence microscopy can be reliably performed. The optimum exposure time can be determined from the exposure reference value according to the method. This allows the photographer to concentrate on other photographing operations other than determining the exposure time, such as determining the angle of view and focusing, and can obtain a higher quality sample photograph. In addition, by using the boundary variance value set based on the variance of each luminance histogram by a microscopic method such as a bright field microscopic method or a fluorescent microscopic method, it is possible to perform highly accurate microscopic discrimination. it can.

In the above-described embodiment, the boundary variance Th is fixed, but a configuration in which the boundary variance Th can be arbitrarily set may be considered. For example, the RAM 13 connected to the CPU 1 is electrically backed up, or a backup memory such as a flash memory or NVRAM is connected, and the boundary variance value T input from the switch input circuit 7 is input.
h may be stored. This is effective in cases where the boundary variance values are offset, instead of general bright-field microscopy and fluorescence microscopy. (Second Embodiment) FIG. 6 shows a schematic configuration of a second embodiment of the present invention, and the same parts as those in FIG. 1 are denoted by the same reference numerals.

In this case, the CPU 1 includes a buzzer circuit 14
Are connected. When the variance of the luminance histogram according to the output of the image sensor 10 is included in a boundary variance range Rg described later, the buzzer circuit 14 warns the photographer of a continuous sound or an intermittent sound that prompts the photographer to select the microscopic method. It emits as a sound.

The ROM 12 stores a control program of the CPU 1, an exposure reference value and a boundary dispersion range Rg for the bright field microscopy and the fluorescence microscopy in the microscope described with reference to FIG. Here, the boundary variance range Rg is determined to a value having the following range using the above-described boundary variance value Th.

Rg (min) ≦ Rg ≦ Rg (max) where Rg (min) = Th−α, Rg (max) = T
h + α and α are arbitrary positive numbers.

Then, the CPU 1 controls the image sensor 10
The illuminance L is obtained from the digital voltage signal corresponding to the current output of the above, the variance of the luminance histogram is obtained, and the microscopic method is determined based on a comparison between the variance of the luminance histogram and the boundary variance range Rg stored in the ROM 12. The exposure reference value K1 or K2 is selected, and the exposure time is further calculated. Further, when the illuminance L is included in a boundary dispersion range Rg to be described later, a warning function is issued via the display circuit 8 and the buzzer circuit 14.

The rest is the same as FIG. Next, the operation of the first embodiment configured as described above will be described with reference to the flowchart of FIG.

First, in step 701, it is determined whether or not a release has been made by the switch input circuit 7. If NO here, at step 702 the switch input circuit 7
It is determined whether the O value has been input. If the ISO value has been input, in step 703, the ISO value is stored in the RAM 13, and the process proceeds to step 704. If the ISO value has been input, the process immediately proceeds to step 704.

In step 704, the illuminance L is calculated. In this case, when a part of the luminous flux transmitted through the objective lens of the observation optical system of the microscope (not shown) enters the image sensor 10, the image sensor 10 outputs a current corresponding to the amount of received light and outputs this current output to the timing generator. The data is output to the A / D conversion circuit 11 at the read timing of No. 9, where it is converted into a digital voltage signal and supplied to the CPU 1. Thereby, the CPU 1 obtains the illuminance L from the digital voltage signal corresponding to the output of the image sensor 10. The CPU 1 determines in step 705 that the image sensor 1
From the digital voltage signal corresponding to the output of 0, a luminance histogram and its variance s are obtained.

Next, at step 706, the CPU 1
The variance .sigma.s of the luminance histogram is compared with the boundary variance range Rg stored in the ROM 12, and the variance .sigma.
g is determined.

In this case, Rg (min) ≦ σs ≦ Rg
If (max), in step 707, the buzzer circuit 14
Warns the photographer to select brightfield microscopy or fluorescence microscopy, and waits for the input of the microscopic method in step 708. Thereafter, the photographer inputs the microscopic method. Then, in step 709, the exposure reference value K1 or K2 corresponding to the input microscopic method is selected.

On the other hand, if the variance s is outside the boundary variance range Rg in step 706, it is determined in step 710 whether the variance s is larger than Rg (max). Here, if the variance s> Rg (max) is satisfied, it is determined that the method is the fluorescence microscopy method. In step 711, the exposure reference value K2 is selected and read from the ROM 12, and the variance s <Rg (mi)
If n) holds, bright field microscopy and determination step 7
At 12, the exposure reference value K1 is selected and read.

Next, in step 713, an identification display corresponding to the microscopic method is displayed on the display circuit 8. A display example in this case is the same as that in FIG. 5 described above. Then, step 71
At 4, the CPU 1 reads out the ISO value stored in the RAM 13, and at step 715, calculates the exposure time. In this case, the exposure time T is obtained from the following equation.

T = K / (AL) where K (K1 or K2) is an exposure reference value, and A is an ISO
The value, L, is the illuminance. Then, in step 716, the calculated exposure time T is displayed on the display circuit 8. The display example in this case is the same as that in FIG. 5 described above.

Next, the process returns to step 701, and thereafter, the above-described operation is repeated while performing the release determination and the ISO value input determination. Thereafter, when it is determined in step 701 that the shutter is released by the switch input circuit 7, in step 717, the shutter 3 is opened by the shutter opening / closing drive circuit 2, and in step 718, it is determined that the exposure time has elapsed. Then, the shutter 3 is closed by the shutter opening / closing drive circuit 2, and then step 720
Thus, the film winding motor drive circuit 4 drives the film winding motor 6 of the camera 5 to wind the photographic film.

Therefore, according to this configuration, in accordance with the effect of the first embodiment, a special bright-field spectroscope or a fluorescence spectroscope whose dispersion is different from that of a general bright-field microscopy or fluorescence microscopy is adopted. Even when the mirror method is used, the speculum method can be reliably selected, so that a high-quality sample photograph with the optimal exposure time can be obtained.

[0045]

As described above, according to the present invention, by observing differences in images due to differences in microscopy by paying attention to the variance of the luminance histogram, an observation system that does not cause fading of the fluorescent specimen is provided. Also when used, the discrimination of the microscopy method can be reliably performed, and the optimal exposure time can be determined from the exposure reference value according to the microscopy method. This allows the photographer to concentrate on other photographing operations other than determining the exposure time, such as determining the angle of view and focusing, and can obtain a higher quality sample photograph.

Further, by using a boundary variance value set based on the variance of each luminance histogram by a microscopic method such as a bright field microscopic method or a fluorescent microscopic method, it is possible to determine a highly accurate microscopic method. It can be carried out.

Furthermore, even when a special bright-field spectroscopy or fluorescence microscopy in which the dispersion is different from general bright-field microscopy or fluorescence microscopy is used, the microscopy can be reliably selected. And a high-quality specimen photograph with an optimal exposure time can be obtained.

[Brief description of the drawings]

FIG. 1 is a diagram showing a schematic configuration of a first embodiment of the present invention.

FIG. 2 is a view showing an exposure reference value according to a microscopy method used in the first embodiment.

FIG. 3 is a view showing an example of dispersion of luminance histograms in a bright field microscopy and a fluorescence microscopy used in the first embodiment.

FIG. 4 is a flowchart for explaining the operation of the first embodiment.

FIG. 5 is a diagram illustrating a display example of the display circuit according to the first embodiment.

FIG. 6 is a diagram showing a schematic configuration of a second embodiment of the present invention.

FIG. 7 is a flowchart for explaining the operation of the second embodiment.

FIG. 8 is a diagram showing a histogram of a typical luminance distribution of a bright field image and a fluorescent image.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 ... CPU, 2 ... Shutter open / close drive circuit, 3 ... Shutter, 4 ... Film winding motor drive circuit, 5 ... Camera, 6 ... Film winding motor, 7 ... Switch input circuit, 8 ... Display circuit, 9 ... Timing generator Reference numeral 10: image sensor, 11: A / D conversion circuit, 12: ROM, 13: RAM, 14: buzzer circuit.

Claims (3)

[Claims] 1. A microscope photographing apparatus for photographing a sample image taken through an observation optical system of a microscope, comprising: a light receiving unit for receiving light transmitted through the observation optical system; Storage means for storing the respective exposure reference values; speculative method discriminating means for discriminating a speculum method using the variance of the luminance histogram according to the output of the light receiving means; An exposure time calculating means for calculating an exposure time based on an exposure reference value according to a microscopy method. 2. A speculum discriminating means, based on a comparison result between a boundary variance value set based on a variance of each luminance histogram by a different speculum method and a variance of a luminance histogram according to an output of the light receiving means. The microscope photographing apparatus according to claim 1, wherein a microscopic method is determined. 3. The speculum method discriminating means determines a boundary variance range using a boundary variance value set based on variances of respective luminance histograms obtained by different speculum methods.
2. The microphotographing apparatus according to claim 1, further comprising a warning unit for issuing a warning for selecting a microscopy method when the variance of the luminance histogram according to the output of the light receiving unit is included in the boundary variance range. .