JPH06235866A - Microscopic device - Google Patents

Abstract

PURPOSE:To prevent a fluorescent sample from fading by shortening a time for switching exciting light, or index light. CONSTITUTION:The device is provided with an illuminating light selecting unit 10 for selectively switching the illuminating light with which a sample 6 is irradiated so as to obtain the illuminating light having different wavelength or brightness, index light source units 17 and 31 for projecting the index light capable of varying the color or the brightness in an observation field of the microscope and a means 30 for interlocking the illuminating light switching operation of the illuminating light selecting unit 10 with the operation of adjusting the color or the brightness of the index light from the index light source units 17 and 31.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a microscope apparatus equipped with a device for switching excitation light, a photographic device for taking an observation image, an index light source unit for indicating a photometric position, and the like.

[0002]

2. Description of the Related Art An example of the structure of such a microscope device is shown in FIG. In the microscope apparatus shown in the figure, the illumination light emitted from the illumination apparatus 1 is incident on the excitation apparatus 2. In the excitation device 2, the excitation filter 3 is arranged on the incident side viewed from the lighting device 1 side. The illumination light incident on the excitation device 2 is transmitted through the excitation filter 3, converted into excitation light having a predetermined excitation wavelength, and further incident on the dichroic mirror 4. The excitation light is reflected by the semi-transmissive surface of the dichroic mirror 4 toward the objective lens 5 side, and the sample 6 is illuminated through the objective lens 5. The sample 6 is fluorescently stained in advance, and a predetermined dye emits fluorescence when excitation light is incident on the sample 6. By the way, in the observation of a fluorescent sample, there are cases where the sample is subjected to multiple staining and the excitation light is sequentially switched to observe the fluorescent dye excited by various excitation lights.

FIG. 8 shows a pumping light switching device 10 configured so that pumping lights having different wavelengths can be selected. In the excitation light switching device 10 shown in the figure, three dichroic mirrors 4-1 to 4-3 are linearly arranged, and the transmission wavelengths are respectively provided on the incident surfaces of the dichroic mirrors 4-1 to 4-3 on the illumination device side. Excitation filters 3-1 to 3-3 having different characteristics are arranged. An absorption filter 7- having absorption wavelengths corresponding to the excitation filters 3-1 to 3-3 respectively corresponding to the end faces of the dichroic mirrors 4-1 to 4-3 from which the light beams incident and transmitted from the objective lens 5 side are emitted. 1 to 7-3 are arranged. One set of the dichroic mirror 4, the excitation filter 3, and the absorption filter 7 constitutes one excitation device 2. The excitation light switching device 10 in which the three cubes are integrated is configured to be slidable in the arrangement direction orthogonal to the optical axis (arrow direction in the drawing).

Object light from the sample 6 (hereinafter referred to as a sample image) again enters the objective lens 5, passes through each dichroic mirror 4, passes through the absorption filter 7, and enters the beam splitter 9 in the lens barrel 8. To do. One of the sample images branched by the beam splitter 9 is guided to the eyepiece lens 11 side and can be observed from the eyepiece lens 11.

The other sample image split by the beam splitter 9 is split into a shooting optical path and a photometric optical path by a beam splitter 12 which is inserted into and removed from the optical path depending on the presence or absence of shooting. The sample image guided to the photographing optical path is formed on the film surface 14 of the camera, for example, via the shutter 13. Further, the sample image guided to the photometric optical path is the photometric sensor 15
Incident on. The photometric output of the photometric sensor 15 is input to an exposure control unit (not shown), and the shutter 13 is opened / closed based on the exposure time calculated there.

Beam splitter 12 and photometric sensor 1
An index member 16 having a pinhole for indicating the position and range of the spot index, and a slotted mirror 17 that can be inserted into and removed from the optical path are arranged on the optical path between the optical path and the optical path.

The index light from the spot light source 18 is incident on the edge mirror 17 disposed on the optical path. When the photometric position in the observation visual field is indicated by the spot index, the Hanenoke mirror 17 is arranged on the optical path, and the index light from the spot light source 18 is incident on the prism 9 via the Hanenoke mirror 17. The index light that has entered the prism 9 is once reflected and then enters the folding mirror 19, where it is reflected, re-enters the prism 9, is combined with the sample image, and is guided to the eyepiece lens 11. As a result, in the observation field of view observed from the eyepiece lens 11, the spot index is superimposed and projected on the sample image.

By the way, the color or brightness of the spot index must be changed according to the wavelength or intensity of the excitation light. Therefore, a plurality of spot light sources 18 with different emission colors are
It has been constructed such that it can be inserted into and removed from the optical axis, and a filter switching device for selectively arranging filters having different transmission wavelengths is arranged on the optical axis side of the spot light source 18.

[0009]

However, in the above-described conventional microscope apparatus, the excitation light switching device 10 is used when an automatic exposure is performed on a sample in which a difference in color or a specific light and shade is generated in the sample image due to the difference in excitation light. Therefore, every time the excitation light is switched, the observer must switch the color or brightness of the spot index by another operation, which is complicated.

When photographing a sample that emits a substantially constant color or brightness for the same illumination intensity, the color or brightness of the spot index may be switched by another operation each time the excitation light is switched. Since the switching operation takes a long time, there is a problem that fading of the fluorescent sample progresses and it becomes difficult to observe under the same conditions.

Further, with the switching of the excitation light, it is necessary to switch the exposure correction value used when calculating the exposure time, but since the photographer also switched the switching operation by another operation, the switching operation takes time. In short, there is a problem that the fluorescent specimen is discolored.

Moreover, when the sample is different, the amount of change in changing the exposure correction value according to the change of the excitation light is also different. Therefore, the operation of changing the exposure correction value becomes more complicated and the change time can be lengthened. It is highly likely.

The present invention has been made in view of the above circumstances, and the switching operation of the excitation light and the adjustment operation of the color or brightness of the index light can be linked, and the exposure correction value can be switched. A microscope device that can be automatically adjusted in association with the adjustment operation of the index light, can speed up the operation related to switching the excitation light or the index light, can simplify the operation, and can prevent the fading of the fluorescent sample. The purpose is to provide.

[0014]

In order to achieve the above object, the present invention corresponding to claim 1 is an illumination light for selectively switching illumination light with which a sample is irradiated to illumination light having different wavelengths or brightness. A selection unit and an index light source unit for projecting index light whose color or brightness is variable within the observation field of view of the microscope,
A configuration is provided that includes means for interlocking the illumination light switching operation of the illumination light selection unit and the index light color or brightness adjustment operation of the index light source unit. The present invention corresponding to claim 2 is a photographing unit for photographing an observation image of a specimen observed by a microscope, and an illumination light for selectively switching the illumination light with which the specimen is irradiated to the illumination light with different wavelength or brightness. A selection unit, an index light source unit that projects index light whose color or brightness is variable in the observation field of view, and a photometric unit that measures the range of at least one pixel indicated on the observation image by the index light, Exposure control means for calculating the exposure time of the photographing unit from the photometric value output from the photometric means and at least the exposure correction value according to the illumination light, and the operation of adjusting the color or brightness of the index light of the index light source unit. And a correction value automatic switching means for switching the exposure correction value of the exposure control means in conjunction with the above.

The present invention corresponding to claim 3 is a photographing unit for photographing an observation image of a specimen observed by a microscope,
An illumination light selection unit that selectively switches the illumination light with which the sample is irradiated to illumination light having a different wavelength or brightness; an index light source unit that projects index light whose color or brightness is variable within an observation field of view; Photometric means for photometrically measuring a range consisting of at least one pixel indicated on the observed image by the index light, lighting control means for lighting the index light source unit at a frequency corresponding to each color of the index light, and output of the photometric means From the indicator light source unit, the indicator light color detecting means for detecting the lighting frequency of the indicator light source unit is provided.

[0016]

In the present invention corresponding to claim 1, the illumination light selecting unit illuminates the sample with predetermined excitation light, and the indicator light source unit responds to the excitation light on the observation image of the sample illuminated by the illumination light. A color or brightness index image is projected. Then, when the illumination light applied to the sample is switched by the illumination light selection unit, the index light projected by the index light source unit in the observation visual field corresponds to the excitation light in conjunction with the switching operation. Adjusted. Alternatively, when the color or brightness of the index light projected by the index light source unit in the observation field of view of the microscope is adjusted, the excitation light of the illumination light selection unit is also switched in association with the operation.

According to the second aspect of the present invention, the illumination light selection unit illuminates the sample with a predetermined excitation light, and the color or brightness index image corresponding to the excitation light from the index light source unit is observed on the observed image of the sample. Is projected. The portion designated by the index image is measured by the photometric means, and the photometric value is given to the exposure control means to calculate the exposure time. The photographing unit is controlled by the exposure time calculated by the exposure control means. When the color or brightness of the index light is adjusted, the exposure correction value of the exposure control means is switched by the correction value automatic switching means in conjunction with the adjustment operation.

In the present invention corresponding to claim 3, the sample is illuminated with predetermined excitation light by the illumination light selection unit, and the color or brightness index image corresponding to the excitation light from the index light source unit is displayed on the observed image of the sample. Is projected. When the color of the index light changes, the index light source unit is turned on by the lighting control means at the frequency corresponding to the changed color of the index light.

On the other hand, the index image projected on the observed image is measured by the photometric means, and the lighting frequency of the index light source unit is detected by the index light color detection means which receives the photometric output, and the color of the changed index light is determined. To be judged.

[0020]

EXAMPLES Examples of the present invention will be described below.

FIG. 1 shows the configuration of a microscope apparatus according to an embodiment of the present invention. The parts having the same functions as those of the device shown in FIG.

In the microscope apparatus of this embodiment, two guides 21 and 22 are arranged in a direction parallel to the sliding direction (direction orthogonal to the optical axis) of the excitation light switching device 10 capable of switching the excitation light. A holding table 23 is slidably supported by the guides 21 and 22. The excitation light switching device 10 is fastened to one surface of the holding table 23, and the rack 22 is arranged on the opposing surface in the sliding direction of the excitation light switching device 10. A motor 25 is arranged near the holding table 23, and a pinion 26 coaxial with the rotation shaft of the motor 25 meshes with the rack 22. A position sensor 27 is provided at a location facing the excitation light switching device 10. The motor 25 is driven and controlled by an excitation light switching control signal provided from the control device 30. In addition, the index light source 3
1 is configured so that the emission color or brightness can be adjusted by the control device 30 and the external switch 32.

FIG. 2 is a diagram showing various operation switches in the control device 30. Each operation switch is contained in the switch board 40. 41-43 are excitation light switching devices 1
A switch for selecting each pumping device 2 of 0, and three types of pumping devices 2 corresponding to the pumping light switching device 10 shown in FIG.
Is selectable. Reference numerals 44 and 45 are switches for selecting the color of the spot index, and in this case, two colors can be selected. Reference numeral 46 denotes a switch for switching the brightness of the spot index, which moves to the left in the figure to be dark and to the right to be bright. Reference numerals 47 to 49 are switches for selecting the amount of change when the exposure correction value is automatically switched, and can be adjusted in three steps.

In order to adjust the color of the spot index (index light), a plurality of index light sources having different emission colors are held so that they can be inserted into and removed from the optical axis, and the index light source is selectively driven electrically. It is realized by being configured to be insertable into the shaft. Alternatively, as shown in FIG. 6, color filters 51 to 53 of a plurality of colors are held on the optical axis side of the index light source 31 so that they can be inserted into and removed from the optical axis, and the color filters 51 to 53 are electrically driven to selectively emit light. This can be realized by configuring the shaft to be removable. The brightness of the index light can be adjusted with the same configuration. The brightness can also be adjusted by controlling the current supplied to the index light source 31. In this embodiment, it is assumed that either configuration is adopted. Next, the operation of this embodiment configured as described above will be described.

In the pumping light switching device 10, one pumping device 2 is selectively arranged on the optical axis from the three pumping devices 2 either manually from the outside or by the control device 30. The processing shown in FIG. 3 is executed in conjunction with the switching operation of the switching device 10. That is, when the excitation light switching device 10 is manually slid from the outside, the position of the excitation light switching device 10, that is, the excitation device 2 inserted on the optical axis is determined from the sensor output of the position sensor 27. Then, the emission color or brightness of the index light source 31 is adjusted to the color or brightness corresponding to the determined excitation light of the excitation device 2. The color or brightness of the index light is switched according to the above-mentioned configuration. The same applies hereinafter.

Further, the operation switches 41 to 41 in the control device 30
When any one of 43 is selected, the current position of the excitation device 2 corresponding to the selected operation switch is determined from the output of the position sensor, and the moving direction and the moving distance of the excitation light switching device 10 are obtained. Then, the drive control of the motor 25 is performed, and the excitation light switching device 10 is moved by the moving distance in the obtained moving direction.
To move. The color or brightness of the index light source 31 is adjusted in association with the movement of the excitation light switching device 10.

As described above, when the pumping light switching device 10 is moved, that is, the pumping light is switched, the control device 30 automatically executes the sequence processing shown in FIG. The color or brightness of 31 is automatically adjusted to the index light corresponding to the switched excitation light.

Further, the indicator light source 31 can switch the emission color or the brightness by the above-mentioned configuration according to an instruction from the external switch 32 or the control device 30. If the switching of the index light was before the switching of the excitation light, the control device 30 operates as shown in FIG.
The process shown in is executed.

When an instruction of the color or brightness of the index light is given from the external switch 32 to the index light source 31, the instruction content of the color or brightness is detected by the sensor 33 and input to the control device 30. The control device 30 detects the color or brightness of the index light switched from the input data, drives the motor 25 according to the detection result, and arranges the excitation device 2 for the corresponding excitation light on the optical axis. . On the other hand, the control device 3
When the color of the index light is selected by any one of the 0 operation switches 44 and 45, the index light source 3 has the configuration described above.
The excitation light switching device 10 operates via the motor 25 so that the excitation device 2 corresponding to the adjusted index light color is arranged on the optical axis while controlling 1 to adjust to the selected color. Controlled. Also, when the brightness of the index light is instructed by the operation switch 46 of the control device 30, the index light source 31 and the excitation device 2 operate in an interlocking manner, as in the case of color selection.

As described above, when there is a switching operation of the color or brightness of the index light, the excitation device 2 is operated so as to be the excitation light corresponding to the color or brightness of the switched index light in conjunction with the operation. Automatically adjust the device. Further, FIG. 5 shows a sequence process in which the exposure correction value and the selection of the color or brightness of the index light are linked.

When an instruction of the color or brightness of the index light is given from the external switch 32 to the index light source 31, the instruction content of the color or brightness is detected by the sensor 33 and input to the control device 30. The control device 30 detects the color or brightness of the index light and switches to an exposure correction value corresponding to the detected color or brightness of the index light. Alternatively, when the color of the index light is selected by one of the operation switches 44 and 45 of the control device 30, the exposure correction value corresponding to the color or the brightness of the selected index light is switched. And the photometric sensor 15
When the exposure control is performed based on the photometric value from, the exposure time is calculated using the exposure correction value set as described above. The shutter 13 is released according to the calculated exposure time.
Is controlled to open and close.

When the exposure correction value is switched according to the index light, the amount of change is the operation switch 4 of the control device 30.
7 to 49 can be selected from three types. Since the switching width of the exposure correction value varies depending on the sample, one of the three switching widths is selected by the operation switches 47 to 49.

As described above, when there is a switching operation of the color or brightness of the index light, the exposure correction value corresponding to the color or brightness of the switched index light is automatically switched in association with the operation. Moreover, the switching width can be arbitrarily set in advance.

As described above, according to the present embodiment, the color or brightness of the index light is automatically adjusted in association with the switching operation of the excitation light, or the excitation or color is excited in association with the operation of switching the color or brightness of the index light. Since the light is automatically adjusted, the switching operation can be simplified and the operation time can be shortened. As a result, unnecessary illumination time can be shortened and fading of the specimen can be prevented.

Further, since the exposure correction value is automatically switched in association with the switching operation of the color or brightness of the index light, unnecessary illumination time can be shortened and fading of the sample can be prevented as in the above. Moreover, the range for changing the exposure correction value is the operation switch 47.
Since a plurality of selections can be made by selecting .about.49, it is possible to interlock the switching of the exposure correction value for many kinds of samples. Although the switch 33 detects the switching of the index light by the external switch 32 in the above embodiment, the same function can be realized by the following configuration.

The index light source 31 is constructed so that lighting can be controlled at a lighting frequency corresponding to the color or brightness of the spot index.
For example, if the spot index is red, the lighting frequency is AH
z, and BHz for green. And the control device 3
0 detects the lighting frequency from the photometric output of the photometric sensor 15 and determines the color of the spot index. With such a configuration, the sensor 33 attached to the external switch 32 can be eliminated. The present invention is not limited to the above embodiments, and various modifications can be made without departing from the gist of the present invention.

[0037]

As described above in detail, according to the present invention, the switching operation of the excitation light and the adjustment operation of the color or brightness of the index light can be linked, and the switching of the exposure correction value can be performed by the index light. Can be automatically adjusted in conjunction with the adjustment operation of
It is possible to provide a microscope apparatus capable of speeding up the operation related to switching of excitation light or index light, simplifying the operation, and preventing fading of the fluorescent sample.

[Brief description of drawings]

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

FIG. 2 is a schematic diagram showing operation switches in a control device provided in the microscope device shown in FIG.

FIG. 3 is a diagram showing a flow of processing from a switching operation of excitation light to a switching operation of color or brightness of index light.

FIG. 4 is a diagram showing a processing flow from a switching operation of the color or brightness of the index light to a switching operation of the excitation light.

FIG. 5 is a diagram showing a flow of processing from an operation of switching the color or brightness of the index light to an operation of switching the exposure correction value.

FIG. 6 is a diagram showing a configuration for switching the color of index light.

FIG. 7 is a configuration diagram of a conventional microscope apparatus.

FIG. 8 is a configuration diagram of an excitation device.

[Explanation of symbols]

DESCRIPTION OF SYMBOLS 1 ... Illumination device, 2 ... Excitation device, 6 ... Sample, 10 ... Excitation light switching device, 13 ... Shutter, 15 ... Photometric sensor, 1
6 ... Index member, 24 ... Rack, 25 ... Motor, 26 ... Pinion, 27 ... Position detection sensor, 30 ... Control device, 31
... Index light source, 32 ... External switch, 33 ... Sensor.

Claims (3)

[Claims] 1. An illumination light selection unit for selectively switching the illumination light with which the sample is irradiated to illumination light having different wavelengths or brightness, and projecting index light with variable color or brightness in the observation field of view of the microscope. A microscope apparatus comprising: an index light source unit; and means for interlocking an illumination light switching operation of the illumination light selection unit with an operation of adjusting the color or brightness of the index light of the index light source unit. 2. A photographing unit for photographing an observation image of a specimen observed by a microscope, an illumination light selection unit for selectively switching the illumination light with which the specimen is irradiated to the illumination light having different wavelengths or brightness, and observation. An index light source unit that projects index light whose color or brightness is variable in the field of view, a photometric unit that measures the range of at least one pixel or more indicated on the observation image by the index light, and an output from the photometric unit Exposure control means for calculating the exposure time of the photographing unit from the photometric value and at least the exposure correction value corresponding to the illumination light, and the operation for adjusting the color or brightness of the index light of the index light source unit. A microscope apparatus comprising: a correction value automatic switching means for switching the exposure correction value of the exposure control means. 3. A photographing unit for photographing an observation image of a specimen observed by a microscope, an illumination light selection unit for selectively switching illumination light with which the specimen is irradiated to illumination light having different wavelengths or brightness, and observation. The index light source unit that projects index light whose color or brightness is variable in the field of view, the photometric unit that measures the range of at least one pixel indicated on the observation image by the index light, and the index light source unit A microscope apparatus comprising: lighting control means for lighting at a frequency corresponding to each color of the index light; and index light color detection means for detecting the lighting frequency of the index light source unit from the output of the photometric means.