JPH05165079A - Microscopic photographing device - Google Patents

Abstract

PURPOSE:To generate a photograph with proper exposure in the photographing system using fluorescent pigments of all light-emission wave-lengths. CONSTITUTION:A microscope photographing device decides the exposure time on the basis of film information and a light measuring signal, wherein a plurality of beams of light having different wave-length ranges are taken out by wave length selecting means 8, 13 selectively from beams of light from a specimen, converted into emitted color component signals, and stored in a first memory means 14, and an emitted wave-length sensing means 15 senses the wave- length emitted by the specimen 1 from the emitted color component signals in the stored wave-length ranges, and a correcting means 16 selects the exposure time corrective value corresponding to the emitted wave-length from a second memory means 17 and performs correction of the exposure time.

Description

Detailed Description of the Invention

[0001]

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

[0002]

2. Description of the Related Art Demand for fluorescence observation is growing not only in the biological field but also in the industrial field. Particularly in the field of biology, due to the nature of the fluorescent specimen to be observed, the specimen will fade over time, so there is a strong demand for obtaining a photograph of proper exposure with one shot of the specimen.

However, fluorescent specimens are not only fading, but are much darker than general bright-field specimens, and in many cases bright spots are extremely present against the background of the darkness.
Since the bright spot is a monochromatic light emission having a peak at the emission wavelength, much higher experience and skill were required to obtain appropriate exposure compared to general bright field specimens.

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

Further, as described in JP-A-57-45514 and JP-B-42-12542,
In order to reliably control the exposure of a dark sample image, a so-called memory metering method has been considered in which the total amount of light is projected onto the film surface after the metering exposure calculation.

[0006]

However, in the photometric method described in each of the above documents, the exposure control is performed in the same manner as in other microscopic observations even when taking a fluorescent photograph, so that the illuminance in which the film exhibits a low illuminance failure characteristic. In the region, the actual density of the film was too high or low, and it was not possible to shoot with proper exposure.

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

The present invention has been made in view of the above circumstances, and it is possible to reliably detect a proper exposure even for a sample image having a characteristic color having a peak at an emission wavelength, and It is an object of the present invention to provide a microscopic photography device capable of obtaining a photograph with various exposures.

[0009]

In order to achieve the above object, the present invention receives a light beam from a sample to be photographed by a light receiving element and converts it into a photometric signal, and film information input from the outside. The exposure time is determined based on the photometric signal, and in the microscopic photography device that controls the opening and closing of the shutter according to the exposure time, the light beam from the sample is incident, and a plurality of light beams having different wavelength regions from the light beam are incident. Wavelength selection means for selectively taking out and converting each to a light emission color component signal, first storage means for storing each light emission color component signal output from the wavelength selection means, and stored in the first storage means From the emitted color component signals of each wavelength range,
The emission wavelength detecting means for detecting the emission wavelength of the sample according to the level state of each emission color component signal, the second storage means for storing the exposure time correction value for each of a plurality of types of emission wavelengths, and the emission An exposure time correction value corresponding to the emission wavelength detected by the wavelength detection means is retrieved from the stored contents of the second storage means, and a correction means for correcting the exposure time is provided.

In order to achieve the above object, a fluorescent filter having a wavelength transmission characteristic corresponding to the fluorescence emission color of the fluorescent sample is arranged on the photographing optical path to form a fluorescent sample image on the film surface, and The light from the fluorescent sample is received by the light receiving element and converted into a photometric signal, the exposure time is determined based on the film information and the photometric signal input from the outside, and the shutter opening / closing control according to the exposure time. In the microscopic photography apparatus for performing, a filter determination unit that determines the type of the fluorescent filter, a storage unit that stores an exposure time correction value according to the type of the fluorescent filter,
And a correction unit that corrects the exposure time by retrieving the exposure time correction value according to the type of the fluorescence filter determined by the filter determination unit from the storage unit.

[0011]

According to the present invention in which the above means are taken, the light rays from the sample are made incident on the wavelength selecting means, and a plurality of light rays having different wavelength regions are selectively taken out into the respective emission color component signals. To be converted. Each of these emission color component signals is stored in the first storage means. Then, the emission wavelength detecting means detects the emission wavelength of the sample from the emission color component signals of each wavelength region stored in the first storage means according to 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.

Further, according to the present invention, the type of the fluorescent filter currently used is detected by the filter discrimination means, and the exposure time correction value corresponding to the detected type of the fluorescent filter is detected from the storage means by the correcting means. Is searched, and the exposure time is corrected by the searched exposure time correction value.

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

[0015]

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

The microscopic photographing apparatus of this embodiment is the same as the sample 1
The sample image is captured by the objective lens 2 which is arranged opposite to, and enters the prism 4 through the photographic lens 3 on which the sample image is formed. This prism 4 collects the incident sample image
It has a semi-transmissive surface 4a divided into a photographing side and a photometric side, and a sample image transmitted through this semi-transmissive surface 4a is incident on a film surface 6 of a camera via a shutter 5. The film surface 6 is placed at the image forming position of the sample 1. Further, the sample image reflected by the semi-transmissive surface 4 a of the prism 4 is incident on the wavelength selection device 8 as the wavelength selection means via the reduction lens 7.

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

The light rays in the respective wavelength regions selectively extracted by the wavelength selecting device 8 are projected on the light receiving element 13. The light-receiving element 13 photoelectrically converts the light beam in each wavelength region by a drive pulse externally applied 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 referred to as “emission color component signal”). Call each) first
It is stored in the memory 4 as a storage means.

The emission color component signal stored in the memory 4 is read 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 utilizes the fact that the emission color component signal of the light beam captured through the filters 10b to 10d having the highest transmittance for the single-wavelength fluorescence emission color of the fluorescence sample has the highest signal level. ing.

The emission wavelength information detected by the emission wavelength detecting means 15 is given to the shutter control section 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 the exposure time correction value for each emission wavelength. In addition, the film sensitivity from the film information input unit 18 to the shutter control unit 16,
Film information such as reciprocity rule is entered.

The shutter control section 16 inputs the photometric signal taken in from the light receiving element 13 by arranging the transparent filter 10a on the optical axis, and calculates the exposure time from the photometric signal and the film information by the exposure time calculation function. To do. Further, in the shutter control unit 16, in the case of the memory photometry method, the emission wavelength detection unit 1 is operated by the function as the correction unit.
The exposure time correction value corresponding to the emission wavelength is read from the ROM 17 on the basis of the emission wavelength information given from 5,
Correct the exposure time. In the case of the real time photometry method, the exposure amount is calculated from the corrected exposure time.

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

In the case of the real-time photometry system, when the release signal is input, the emission wavelength is detected. A timing signal is given from the shutter controller 16 to the motor control means 12, and the motor control means 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 captured through the transparent filter 10a is input to the shutter control section 16 where the illuminance of the sample image is detected and the exposure time is calculated from the illuminance information and the filter information. ..

Light rays captured through the filters 10b to 10d are sequentially photoelectrically converted by the light receiving element 13 and stored in the memory 14. The emission wavelength detecting means 15
The emission wavelength is detected by and is provided to the shutter controller 16.

In the shutter control section 16, the ROM is based on the emission wavelength information given from the emission wavelength detecting means 15.
The exposure time correction value corresponding to the emission wavelength is read from 17. Then, the calculated exposure time is corrected using this exposure time correction value, and the 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 at the same time when the photographing is started, and the photometric signal output from the light receiving element 13 is taken in to start the photometry. Then, the photometric signal is sequentially calculated to detect the exposure amount in real time, and when the calculated exposure amount is reached, the shutter 5 is closed.

Further, FIG. 2B shows a flow chart in the case of operating by the memory photometry method. In this case, the exposure time correction value is selected from the contents stored in the ROM 17 in the same manner as described above, and the exposure time is corrected in the same manner as described above. Then, the shutter 5 is opened for shooting for the corrected exposure time.

As described above, according to this embodiment, the sample image is converted into the red transmission filter 10b, the green transmission filter 10c,
The emission wavelength is detected by capturing it through the blue transmission filter 10d, and the exposure time correction value corresponding to the detected emission wavelength is retrieved from the ROM 17, which stores the exposure time correction value for each emission color, and Since the exposure time calculated from the photometric signal is corrected, even when taking a photograph of a sample that has a characteristic color that has a peak at its emission wavelength, such as a fluorescent sample, it is extremely easy to optimize It can be an exposure and you can get a photo of the proper exposure. Next, a second embodiment of the present invention will be described. FIG. 3 shows the configuration of the microscope photography apparatus according to the second embodiment. The parts having the same functions as those of the device shown in FIG. 1 are designated 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. The other structure is similar to that of the first embodiment. That is,
The color sensor 20 realizes a function as a wavelength selection unit and a function as a light receiving element.

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, and the light transmitted through each of the filters 21a, 21b, and 21c is photoelectrically converted and output to the memory 14. Is becoming The reference numerals 22 and 23 shown in FIG.
Is driven for each of the R, G, and B filters to individually take out the 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 generate a photometric signal. Can be created.

In this embodiment, the color sensor 20 is used.
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 after being photoelectrically converted via 21b and 21c.
Further, the photometric signal output from the color sensor 20 is input to the shutter controller 16. Then, the exposure time is corrected in the same manner as described below, and a photograph is taken.

According to the present embodiment, the color sensor 20 takes in the emission color component signals in the respective wavelength regions of R, G and B, so that the time for rotating the filter turret 9 is longer than that in the first embodiment. , Which is more effective in avoiding the fading of fluorescent specimens. Further, since the wavelength selection device 8 for taking in the emission color component signal of each wavelength region can be removed from the device, 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 the microscope photography apparatus according to the third embodiment. The parts having the same functions as those of the device shown in FIG. 1 are designated by the same reference numerals.

In the microscope photography apparatus of this embodiment, a fluorescence filter set 30 is arranged on the optical path between the photography 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 fluorescence filter set 30 via the collector lens 33.

Further, the filter discriminating means 3 for detecting the kind of the fluorescent filter set 30 arranged on the optical path.
4 are provided. The type determination signal of the fluorescence filter set 30 detected by the filter determination means 34 is input to the shutter controller 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 fluorescence emission color exhibited by the fluorescence sample image is determined to some extent depending on the type of fluorescence filter used. It is known that there is a relationship shown in FIG. 6 between the type of the fluorescent filter and the fluorescence 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. In practice, the fluorescent emission color portions of the storage table are replaced with the corresponding emission wavelengths.

The shutter control section 35 has a function as a correction means, and retrieves the corresponding exposure time correction value from the ROM 36 based on the discrimination signal given from the filter discrimination means 34.

In the present embodiment configured as described above, the type of the fluorescence filter set 30 is detected by the filter discrimination means 34 and given to the shutter controller 35, and the corresponding exposure from the ROM 36 is performed based on the discrimination signal. The time correction value is selected. Therefore, according to this embodiment, the same effect as that of the first embodiment can be obtained.

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

[0043]

As described in detail above, according to the present invention, 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, and to obtain an appropriate exposure. It is possible to provide a microscope photography apparatus capable of obtaining a photograph.

[Brief description of drawings]

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

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

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

FIG. 4 is a perspective view of a color sensor included in the microscope photography apparatus of the second embodiment.

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

FIG. 6 is an RO equipped in the microscope photography apparatus of the third embodiment.
The figure which shows a part of memory content of M.

[Explanation of symbols]

1 ... specimen, 2 ... objective lens, 3 ... photography lens, 4 ...
Prism, 5 ... Shutter, 6 ... Film surface, 8 ... Wavelength selection device, 13 ... Light receiving element, 14 ... Memory, 15 ... Emission wavelength detecting means, 16 ... Shutter control section, 17 ... RO
M, 18 ... Film information input means.

Claims (3)

[Claims] 1. A light receiving element receives a light beam from a sample to be photographed and converts it into a photometric signal, and determines an exposure time based on the film information and the photometric signal input from the outside, and the exposure time is determined. In a microscopic photography device that controls opening and closing of a shutter according to time, a light beam from the sample is incident, a plurality of light beams having different wavelength regions are selectively extracted from the light beam, and wavelength selection is performed to convert each light emission color component signal. Means, first storage means for respectively storing each emission color component signal output from the wavelength selection means, and each emission from the emission color component signals of each wavelength region stored in the first storage means. An emission wavelength detecting means for detecting an emission wavelength of the sample according to a level state of a color component signal; a second storage means storing an exposure time correction value for each of a plurality of types of emission wavelengths; A micrograph comprising: a correction unit that searches the stored contents of the second storage unit for an exposure time correction value corresponding to the emission wavelength detected by the wavelength detection unit and corrects the exposure time. Imaging device. 2. The microscope image taking apparatus according to claim 1, wherein the wavelength selecting means is composed of a color area sensor. 3. A fluorescent filter having a wavelength transmission characteristic according to the fluorescent emission color of the fluorescent sample is arranged on the photographing optical path to form an image of the fluorescent sample on the film surface, and to receive light rays from the fluorescent sample. In the photomicrographing 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, Filter discrimination means for discriminating the type of the fluorescent filter, storage means for storing the exposure time correction value according to the type of the fluorescent filter, and depending on the type of the fluorescent filter discriminated by the filter discriminating means And a correction unit that corrects the exposure time by retrieving an exposure time correction value from the storage unit.