JP3022670B2 - Microscope photography equipment - Google Patents

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 specimen image observed under a microscope.

[0002]

2. Description of the Related Art The demand for fluorescence observation is growing not only in the biological field but also in the industrial field. In particular, in the field of living organisms, due to the nature of the fluorescent specimen to be observed, the specimen fades with the passage of time, and there is a high demand to surely obtain a properly exposed photograph by one specimen photographing.

However, there are many fluorescent specimens which are not only faded but also very dark as compared with general bright-field specimens, and have bright spots extremely against the dark background.
Since the luminescent spot is a monochromatic emission having a peak at the emission wavelength, much more advanced experience and skill were required in order to obtain proper exposure as compared with a general bright-field specimen.

Therefore, a so-called real-time photometric microscope photographing apparatus which is not affected by a change in light amount due to fading of a specimen is disclosed in Japanese Patent Application Laid-Open No. 56-52730 by the present application.

As described in JP-A-57-45514 and JP-B-42-12542,
In order to reliably control the exposure of a dark specimen image, a so-called stored photometry method has been considered in which the total light quantity is projected onto a film surface after photometric exposure calculation.

[0006]

However, in the photometric method described in each of the above-mentioned documents, the exposure control is performed in the same manner as in the case of other microscopy when taking a fluorescent photograph, so that the illuminance of the film showing a low illuminance failure characteristic. In the area, the film exposure density was excessive or insufficient, and it was not possible to take a picture with proper exposure.

In particular, during fluorescence observation, the fluorescent specimen image exhibits a characteristic color having a peak at its emission wavelength,
When a color film is used, the R, G, and B emulsion layers cause low illuminance failure at different ratios, and there is a problem that a photograph with proper exposure cannot be obtained depending on the color of the fluorescent dye used.

SUMMARY OF THE INVENTION The present invention has been made in view of the above circumstances, and it is possible to reliably detect an appropriate exposure even for a sample image having a characteristic color having a peak in the emission wavelength. It is an object of the present invention to provide a microphotographing device capable of obtaining a photograph with a high exposure.

[0009]

According to the present invention, in order to attain the above object, a light beam from a specimen to be photographed is received by a light receiving element, converted into a photometric signal, and film information input from the outside is obtained. An exposure time is determined based on the photometric signal, and in a photomicrographing apparatus that controls opening and closing of a shutter according to the exposure time, a light beam from the sample enters, and a plurality of light beams having different wavelength ranges from the light beam. Wavelength selecting means for selectively taking out and converting each to a luminescent color component signal, first storing means for storing each luminescent color component signal output from the wavelength selecting means, and storing in the first storing means, respectively. From the emission color component signals of each wavelength region
Emission wavelength detection means for detecting the emission wavelength of the sample in accordance with the level state of each of the emission color component signals; second storage means for storing an exposure time correction value for each of a plurality of types of emission wavelengths; Correction means for correcting the exposure time by retrieving an exposure time correction value corresponding to the emission wavelength detected by the wavelength detection means from the contents stored in the second storage means.

In order to achieve the above object, a fluorescent filter having a wavelength transmission characteristic corresponding to a fluorescent emission color of a fluorescent sample is arranged on a photographing optical path to form an image of the fluorescent sample on a film surface; The light from the fluorescent specimen is received by a light receiving element and converted into a photometric signal, an exposure time is determined based on film information input from the outside and the photometric signal, and shutter opening / closing control according to the exposure time is performed. In the microphotographing device performing, the filter determination means for determining the type of the fluorescent filter, and storage means in which an exposure time correction value is stored according to the type of the fluorescent filter,
Correction means for searching the storage means for an exposure time correction value corresponding to the type of the fluorescence filter determined by the filter determination means and correcting the exposure time.

[0011]

According to the present invention in which the above means are taken, a light beam from the sample enters the wavelength selection means, where a plurality of light rays having different wavelength ranges are selectively extracted and converted into emission color component signals. Is converted. These emission color component signals are respectively stored in the first storage means. Then, the emission wavelength detection unit detects the emission wavelength of the sample from the emission color component signals of each wavelength region stored in the first storage unit in accordance with the level state of each emission color component signal.

When the emission wavelength of the sample is detected in this way, the correction means retrieves the exposure time correction value corresponding to the detected emission wavelength from the second storage means, and corrects the exposure time.

According to the present invention, the type of the currently used fluorescent filter is detected by the filter determining means, and the exposure time correction value corresponding to the detected type of the fluorescent filter is stored in the storage means by the correcting means. Is retrieved, and the exposure time is corrected by the retrieved exposure time correction value.

In the case of fluorescent observation, the color of the fluorescent light to be observed is determined by the type of fluorescent filter to be used. By detecting the type of fluorescent filter, fluorescent emission color information can be obtained.

[0015]

Embodiments of the present invention will be described below with reference to the drawings. FIG. 1 shows a configuration of a microscope photographing apparatus according to a first embodiment of the present invention.

The microscope photographing apparatus of the present embodiment
The sample image is captured by an objective lens 2 disposed to face the lens, and the sample image is incident on a prism 4 via a photographic lens 3 on which the sample image is formed. The prism 4 converts the incident sample image into
It has a semi-transmissive surface 4a for dividing into a photographing side and a photometric side, and a sample image transmitted through the semi-transmissive surface 4a is incident on a film surface 6 of a camera via a shutter 5. This film surface 6 is located at the image forming position of the specimen 1. The sample image reflected by the semi-transmissive surface 4a of the prism 4 is incident on a wavelength selection device 8 as a wavelength selection unit via a reduction lens 7.

The wavelength selecting device 8 includes a filter turret 9, a transparent filter 10a having the same transmittance over the entire visible light range, a red transmittance filter 10b having a transmittance peak at a wavelength of 575 (nm), and a wavelength 525 (nm). Green transmission filter 10c having a transmittance peak, wavelength 450
Blue transmission filter 10 having transmittance peak at (nm)
d, and the filters 10a to 10d are arranged on the same circumference around the rotation axis so that the filters 10a to 10d are sequentially arranged on the optical axis by rotating the filter turret 9. The rotation axis of the motor 11 is connected to the center axis of the filter turret 9, and the rotation of the motor 11 is controlled by the motor control means 12.

Light beams in each wavelength region selectively extracted by the wavelength selection device 8 are projected onto the light receiving element 13. The light receiving element 13 photoelectrically converts the light beam in each wavelength region by a drive pulse supplied from the outside in synchronization with the rotation of the filter turret 9, and converts the light beam in each wavelength region into a photoelectric conversion signal (hereinafter, “emission color component signal”). Call the first)
Is stored in the memory 4 as storage means.

The emission color component signal stored in the memory 4 is read out from the emission wavelength detecting means 15, and the emission color component signal having the highest signal level is detected as the emission wavelength.
This is based on the fact that the luminescent color component signal of the light beam captured through the filters 10b to 10d having the highest transmittance for the single-wavelength fluorescent emission color of the fluorescent sample has the highest signal level. ing.

The emission wavelength information detected by the emission wavelength detecting means 15 is given to a shutter control unit 16 having a function as a correcting means, an exposure time calculating function, and a function as a shutter driver.

On the other hand, the ROM 17 stores an exposure time correction value for each emission wavelength. In addition, the film information input means 18 sends the film speed,
Film information such as reciprocity irregularity is input.

The shutter control unit 16 arranges the transparent filter 10a on the optical axis, inputs a photometric signal taken in from the light receiving element 13, and calculates an exposure time by an exposure time calculation function from the photometric signal and the film information. I do. Further, in the case of the storage photometry method, the shutter control unit 16 uses the function as a correction unit to operate the emission wavelength detection unit
Reading an exposure time correction value corresponding to the emission wavelength from the ROM 17 based on the emission wavelength information given from 5,
The above exposure time is corrected. In the case of the real-time photometry method, the exposure amount is further calculated from the corrected exposure time.

Next, the operation of the present embodiment configured as described above will be described with reference to FIG. FIG. 2A shows a flowchart in the case of operating by the real-time photometry method.

In the case of the real-time photometry method, when a release signal is input, the emission wavelength is detected. In this case, a timing signal is given from the shutter control unit 16 to the motor control unit 12, and the motor control unit 12 drives the motor 11 based on the timing signal. Then, the transparent filter 10a, the red transmission filter 10b, the green transmission filter 10c, and the blue transmission filter 10d of the filter turret 9 are sequentially arranged on the optical axis.

The photometric signal of the light beam received through the transparent filter 10a is input to the shutter control unit 16, where the illuminance of the sample image is detected, and the exposure time is calculated from the illuminance information and the filter information. .

The light rays taken in through the filters 10b to 10d are sequentially photoelectrically converted by the light receiving element 13 and stored in the memory 14. And the emission wavelength detecting means 15
The light emission wavelength is detected by the control signal and supplied to the shutter control unit 16.

In the shutter control section 16, a ROM based on the emission wavelength information given from the emission wavelength detection means 15 is used.
From 17, an exposure time correction value corresponding to the emission wavelength is read. Then, the calculated exposure time is corrected using the exposure time correction value, and an exposure amount is calculated from the corrected exposure time and the illuminance.

Next, the transparent filter 10a is arranged on the optical axis, the shutter 5 is opened simultaneously with the start of photographing, and the photometry signal output from the light receiving element 13 is taken in to start photometry. Then, the photometric signals are sequentially calculated, the exposure amount is detected in real time, and when the calculated exposure amount is reached, the shutter 5 is closed.

FIG. 2B shows a flowchart in the case of operating by the stored photometry method. In this case, the exposure time correction value is selected from the contents stored in the ROM 17 as described above, and the exposure time is corrected as described above. Then, the shutter 5 is opened for the corrected exposure time to perform photographing.

As described above, according to the present embodiment, the sample image is transformed into the red transmission filter 10b, the green transmission filter 10c,
The emission wavelength is detected through the blue transmission filter 10d, and the exposure time correction value corresponding to the detected emission wavelength is searched from the ROM 17 storing the exposure time correction value for each emission color. The exposure time calculated from the photometric signal is corrected so that even a photograph of a specimen exhibiting a characteristic color having a peak at its emission wavelength, such as a fluorescent specimen, is extremely easily and optimally photographed. Exposure can be obtained, and a photograph with an appropriate exposure can be obtained. Next, a second embodiment of the present invention will be described. FIG. 3 shows the configuration of a microphotographing apparatus according to the second embodiment. Parts having the same functions as those of the apparatus shown in FIG. 1 are denoted by the same reference numerals.

In this embodiment, the wavelength selecting device 8 provided in the first embodiment is removed from the optical system for photometry, and a color sensor 20 is provided as a light receiving element instead. Other configurations are the same as in the first embodiment. That is,
The function as the wavelength selection means and the function as the light receiving element are realized by the color sensor 20.

The color sensor 20, as shown in FIG.
R, G, and B filters 21a, 21b, and 21c are provided on the surface that receives the light beam from the prism 4. Has become. The reference numerals 22 and 23 shown in FIG.
Are driven for each of the R, G, and B filters to individually extract photoelectric conversion signals.

The filters 21a, 21b and 21c function in the same manner as the filters 10b to 10d arranged in the filter turret 9 of the first embodiment, and combine the respective outputs of R, G and B to convert the photometric signal. Can be created.

In the present embodiment, the color sensor 20
The sample image incident on the R, G, B filters 21a,
The light-emission color component signals in the respective wavelength regions are stored in the memory 14 by photoelectric conversion through the respective 21b and 21c.
Further, a photometric signal output from the color sensor 20 is input to the shutter control unit 16. Then, exposure time correction is performed in the same manner, and photographing is performed.

According to the present embodiment, the color sensor 20 takes in the emission color component signals in the R, G, and B wavelength regions, so that the time required for rotating the filter turret 9 is longer than in the first embodiment. This is more effective in avoiding fading of the fluorescent specimen. Further, the wavelength selection device 8 for taking in the emission color component signals of each wavelength region can be eliminated from the device, so that the configuration can be simplified.
Further, if a color area sensor is used as the color sensor 20, it is effective for photographing a sample having a finer distribution. Next, a third embodiment of the present invention will be described. FIG. 5 shows the configuration of a microscope photographing apparatus according to the third embodiment. Parts having the same functions as those of the apparatus shown in FIG. 1 are denoted by the same reference numerals.

In the microphotographing apparatus of this embodiment, a fluorescent filter set 30 is arranged on the optical path between the photographing lens 3 and the prism 4. And the light source lighting device 3
The light generated by turning on the light source 32 by 1 is incident on the fluorescent filter set 30 via the collector lens 33.

A filter discriminating means 3 for detecting the type of the fluorescent filter set 30 arranged on the optical path.
4 are provided. The type discrimination signal of the fluorescent filter set 30 detected by the filter discriminating means 34 is input to the shutter control unit 35. The shutter control unit 35 further receives film information from the film information input unit 18 and a photometric signal from the light receiving element 13.

Here, in the case of fluorescence observation, the fluorescent emission color of the fluorescent specimen image is determined to some extent depending on the type of the fluorescent filter used. It is known that there is a relationship shown in FIG. 6 between the type of fluorescent filter and the fluorescent emission color.

Therefore, in this embodiment, the ROM 36 has a storage table shown in FIG. 6 in addition to the exposure time correction value corresponding to each emission wavelength. Actually, the fluorescent emission color portions in the storage table are replaced with the corresponding emission wavelengths.

The shutter control unit 35 has a function as a correction unit, and searches a corresponding exposure time correction value from the ROM 36 based on a determination signal given from the filter determination unit 34.

In this embodiment constructed as described above, the type of the fluorescent filter set 30 is detected by the filter discriminating means 34 and given to the shutter control unit 35, and the corresponding exposure light is read from the ROM 36 based on the discrimination signal. A time correction value is selected. Therefore, according to the present embodiment, the same effects as those of the first embodiment can be obtained.

Since the present invention obtains the output of the light receiving element for each of a plurality of different wavelengths, a function as a color thermometer capable of measuring the color temperature of a sample is added by adding a simple circuit. Can be done.

[0043]

As described above in detail, according to the present invention, it is possible to reliably detect an appropriate exposure even for a sample image exhibiting a characteristic color having a peak in the emission wavelength, and to obtain an appropriate exposure. Can be provided.

[Brief description of the drawings]

FIG. 1 is a configuration diagram of a microscope photographing apparatus according to a first embodiment of the present invention.

FIG. 2 is a diagram showing operation steps of the microscope photographing apparatus according to the first embodiment.

FIG. 3 is a configuration diagram of a microscope photographing apparatus according to a second embodiment of the present invention.

FIG. 4 is a perspective view of a color sensor provided in a microscope photographing apparatus according to a second embodiment.

FIG. 5 is a configuration diagram of a microscope photographing apparatus according to a third embodiment of the present invention.

FIG. 6 is a diagram showing an RO provided in a microscope photographing apparatus according to a third embodiment.
The figure which shows some storage contents of M.

[Explanation of symbols]

1 ... sample, 2 ... objective lens, 3 ... photography lens, 4 ...
Prism, 5: shutter, 6: film surface, 8: wavelength selecting device, 13: light receiving element, 14: memory, 15: emission wavelength detecting means, 16: shutter control unit, 17: RO
M, 18 ... film information input means.

Claims (3)

(57) [Claims] 1. A light beam from a specimen to be photographed is received by a light receiving element and converted into a photometric signal. An exposure time is determined based on film information input from the outside and the photometric signal. In a microphotographing apparatus that controls opening and closing of a shutter in accordance with time, a light beam from the sample enters, a wavelength selection for selectively extracting a plurality of light beams having different wavelength ranges from the light beam and converting the light beams into emission color component signals. Means; first storage means for storing the respective emission color component signals output from the wavelength selection means; and emission light component signals for each wavelength region stored in the first storage means. Emission wavelength detection means for detecting the emission wavelength of the sample according to the level state of the color component signal; second storage means for storing an exposure time correction value for each of a plurality of types of emission wavelengths; A correction means for retrieving an exposure time correction value corresponding to the emission wavelength detected by the wavelength detection means from the contents stored in the second storage means and correcting the exposure time. Shooting equipment. 2. The apparatus according to claim 1, wherein said wavelength selecting means comprises a color area sensor. 3. A fluorescent filter having a wavelength transmission characteristic according to a fluorescent emission color of a fluorescent sample is arranged on a photographing optical path to form an image of the fluorescent sample on a film surface, and receives a light beam from the fluorescent sample. In a microphotographing device that receives light by the element and converts it into a photometric signal, determines the exposure time based on the film information and the photometric signal input from the outside, and performs shutter opening / closing control according to the exposure time, A filter discriminating unit for discriminating the type of the fluorescent filter; a storage unit storing an exposure time correction value according to the type of the fluorescent filter; and a type corresponding to the type of the fluorescent filter determined by the filter determining unit. A correction means for retrieving an exposure time correction value from the storage means and correcting the exposure time.