JP4503247B2 - Fluorescence microscope - Google Patents

Description

[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a fluorescence microscope.
[0002]
[Prior art]
As a kind of microscope, a fluorescence microscope for observing fluorescence emitted from a sample is known. There are two types of fluorescent microscopes, an upright type in which an objective lens is disposed above the stage and an inverted type in which the objective lens is disposed below the stage. The basic configuration of the fluorescence microscope includes an epi-illumination unit that irradiates the sample with excitation light and a fluorescence observation unit that images and observes the fluorescence emitted from the sample, and the fluorescence observation unit is for observation with the human eye The eyepiece optical system and / or an imaging unit such as a CCD camera, and the excitation light from the epi-illumination unit is applied to the sample through a filter.
[0003]
Patent Document 1 discloses that an arbitrary filter cassette is set by sliding a plurality of filter cassettes arranged in a line in the horizontal direction. Patent Document 1 also discloses that the excitation method and the wavelength characteristic are indicated on each filter cassette so that the characteristics of the set filter cassette can be visually confirmed through this notation.
[0004]
By the way, a fluorescent microscope has been generally installed in a dark room and observed in the dark room. In recent years, an image display device such as a CRT or LCD and / or a personal computer is connected to the fluorescent microscope with a cable. In many cases, a fluorescent image is displayed on a CRT or the like for observation or image data is processed by a personal computer.
[0005]
In such a usage pattern, light emitted from a CRT or the like may enter the fluorescence microscope and cause the quality of the fluorescence image to deteriorate, and in addition, the operation of the personal computer in the dark room is inconvenient. There's a problem.
[0006]
As one means for solving such a problem, Patent Document 2 proposes to provide a stage on which a sample is placed and a wall for shielding light around the objective lens. According to this proposal, there is an advantage that it is not necessary to work in a dark room for fluorescence observation.
[0007]
[Patent Document 1]
JP-A-8-338947 [Patent Document 2]
Japanese Patent Laid-Open No. 2002-207177
[Problems to be solved by the invention]
When a fluorescent microscope can be used even outside a dark room, it is inevitable that a request to use the fluorescent microscope on a private desk in a laboratory where daily work is performed will appear. Miniaturization of the microscope is required. In addition, as in the past, there is naturally a demand for downsizing even in a fluorescent microscope of the type used in a dark room.
[0009]
Accordingly, an object of the present invention is to provide a fluorescent microscope that can be miniaturized.
[0010]
[Means for Solving the Problems]
According to the present invention, the above technical problem is
A stage on which the sample is placed ;
A transmission illumination unit disposed above the stage and irradiating the sample on the stage with transmission illumination;
An objective lens disposed below and adjacent to the stage ;
A filter disposed below and in the vicinity of the objective lens ;
An epi-illumination tube disposed on a first horizontal axis perpendicular to the filter ;
An epi-illuminator attached to an end of the epi-illumination tube and for irradiating the sample on the stage with excitation light ;
An imaging mirror disposed below and in the vicinity of the filter;
An imaging cylinder arranged on a second horizontal axis arranged facing the imaging mirror;
An imaging device attached to an end of the imaging cylinder,
The epi-illumination tube and the imaging tube are arranged in a V shape having an acute angle when the first horizontal axis and the second horizontal axis are viewed from above ,
This is achieved by providing a fluorescent microscope characterized in that the epi-illuminator and the imaging device are arranged at a height position where they interfere with each other when viewed from the side .
[0011]
That is, in the fluorescence microscope according to the present invention, the first horizontal axis of the epi-illumination tube and the second horizontal axis of the imaging tube are arranged in a V shape having an acute angle when viewed from above. Therefore, the epi-illumination unit and the image-capturing unit can be arranged close to each other without worrying about interference between the epi-illuminator and the image-capturing device, thereby enabling miniaturization.
[0012]
Since the epi-illumination unit and the imaging unit can be arranged close to each other, the length of the imaging cylinder constituting a part of the imaging unit, that is, the imaging cylinder arranged between the imaging mirror and the imaging device From this point of view, the fluorescent microscope can be reduced in size.
[0013]
【Example】
1st Example (FIGS. 1-9)
FIG. 1 is a perspective view showing the internal structure of an inverted fluorescence microscope. In the figure, a fluorescence microscope 1 has a stage 2, and a sample 3 is placed on the stage 2. The stage 2 is fixed in the vertical direction as in the prior art, and a Z-axis stage of an objective lens 10 to be described later can move in the vertical direction.
[0014]
The fluorescence microscope 1 includes a transmission illumination unit including a halogen lamp 5 that irradiates the sample 3 with transmission illumination via an illumination mirror 4 disposed above the stage 2 on the side opposite to the side on which the researcher accesses the stage 2. 6 and an epi-illumination unit 8 including a mercury lamp 7 that irradiates excitation light from below the stage 2, and these are selectively used. Note that reference numeral 9 in FIG. 1 is a support base, and the transmitted illumination unit 6 is fixed on the support base 9.
[0015]
An objective lens 10 is provided directly below the stage 2, and a filter cassette 11 is disposed under the objective lens 10. Since the specific configuration of the filter cassette 11 is the same as that of the filter cassette described in Patent Document 1, detailed description thereof is omitted.
[0016]
An epi-illumination unit 8 is disposed in association with the filter cassette 11. The epi-illumination unit 8 includes an epi-illumination tube 12 disposed on a horizontal axis L1 orthogonal to the filter cassette 11. A mercury lamp 7 as an epi-illuminator is attached to the end of the tube 12.
[0017]
An imaging mirror 13 is disposed directly below the filter cassette 11. An imaging cylinder 14 is disposed on the horizontal axis L2 so as to face the imaging mirror 13, and an imaging device (for example, at the end of the imaging cylinder 14). A CCD camera 15) is attached. Reference numeral 16 in FIG. 1 is a power supply board, and 17 is a control board. The imaging data from the CCD camera 15 is supplied to an image display device such as a personal computer and / or a CRT via a cable not shown.
[0018]
The epi-illumination tube 12 and the imaging tube 14 are arranged in a V shape having a sandwiching angle of an angle θ when viewed from above, so that the mercury lamp 7 and the CCD camera 15 are viewed from above. Sometimes it is arranged in a state offset to the left and right.
[0019]
In this embodiment, the filter cassette 11 includes four different types of filter cassettes 11 so that an appropriate filter cassette 11 can be selected by a researcher's manual operation. As best understood from FIGS. 2 and 3, the guide rail 20 extending perpendicular to the horizontal axis L <b> 1 of the epi-illumination tube 12 is provided with a slider 22 slidably fitted in the horizontal groove 21. Four filter cassettes 11 are attached. The slider 22 and the filter cassette 11 are positioned by dovetail fitting, and the filter cassette 11 can be attached to and detached from the slider 22.
[0020]
The slider 22 has a spur gear 24 at its lower end edge, and the spur gear 24 forms a rack and pinion mechanism with a circular gear 25 meshing with the spur gear 24. That is, as the circular gear 25 rotates, the slider 22 slides along the guide rail 20 in the horizontal direction, and thereby the filter cassette 11 moves linearly.
[0021]
The circular gear 25 is fixed to one end of the rotating shaft 26, and an operation dial 27 is attached to the other end of the rotating shaft 26. Further, the rotary shaft 26 is provided with a circular seal member 28 (FIG. 3) between the circular gear 25 and the operation dial 27. In FIG. 1, the circular seal member 28 is omitted in order to avoid complication of the diagram.
[0022]
The circular seal member 28 has flanges 30 at both ends, and a recess 31 is formed between the pair of flanges 30 and 30. The flange 30 is not particularly limited but may be circular.
[0023]
As shown in FIG. 4, the fluorescence microscope 1 having the above-described internal structure has a frame structure in which a plurality of columns 36 are erected on a base 35, and the support 36 for transmitted illumination is supported by the columns 36. The imaging device 14 and the like are built in the base 35.
[0024]
A circular hole (not shown) is provided at the end of the top plate 37 of the base 35, and the filter cassette 11 is moved in a state in which the recess 31 of the circular seal member 28 described above is fitted in this circular hole. A cassette actuating mechanism is provided. That is, the cassette operating mechanism including the operation dial 27, the circular gear 25, the circular seal member 28 and the like described above can be best understood from FIG. 3 in the recess 31 of the circular seal member 28 in the base top plate 37. It arrange | positions in the state which the periphery of the circular hole entered, and the opening edge of this circular hole will be in the state covered with a pair of circular flanges 30 and 30 of a circular sealing member. The circular gear 25 is positioned on the base top surface plate 37, and the operation dial 27 is positioned below the base top surface plate 37. The circular seal member 28 may be rotatable with respect to the rotation shaft 26, or may rotate with the rotation shaft 26 so as to be in sliding contact with the circular hole of the base top surface plate 37.
[0025]
Filter cassette feeding mechanisms such as a plurality of filter cassettes 11 and guide rails 20 are preferably housed in an integrated case 38 as shown in FIG. 6, and a display window 39 is provided in the case 38, and this display window It is further preferable to display the characteristics of the currently set filter cassette 11 through 39.
[0026]
The fluorescent microscope 1 may be used in the state shown in FIG. 4 when used in a dark room. However, when used outside the dark room, the fluorescent microscope 1 is used with a light shielding box 40 as shown in FIG. Is good. The light shielding box 40 has a size capable of covering the other components as a whole with the operation dial 27 exposed to the outside, and is fixed to the base 35 so as to be detachable, for example. The light shielding box 40 covers the entire internal structure of the fluorescent microscope 1 and shields it from light. However, the light shielding box 40 has a door 41 at a position corresponding to the stage 2, and opens and closes the sample 3. Can do. The size of the door 41 is preferably set to such a size that the above-described display window 39 can be seen from the outside when the door 41 is opened.
[0027]
According to the fluorescence microscope 1 of the embodiment, as best understood from FIG. 2 , the epi-illumination tube 12 and the imaging tube 14 are arranged in a V shape when viewed from above, and each tube 12 The mercury lamp 7 and the image pickup device 15 located at the ends of 13 and 13 are offset from each other right and left, so that there is no problem of interference between the mercury lamp 7 and the image pickup device 15 and the epi-illumination tube 12 And the imaging cylinder 14 can be made as close as possible to each other.
[0028]
In other words, as compared with the case of arranging vertically a mercury lamp 7 and the imaging device 15 in the same vertical plane, without having to worry about interference between the mercury lamp 7 and the imaging device 15, located below the filter cassettes 11 Since the imaging mirror 13 to be moved can be disposed close to the filter cassette 11, the height dimension of the fluorescence microscope 1 can be reduced.
[0029]
Further, by arranging the filter cassette 11 and the imaging mirror 13 close to each other, the length of the imaging cylinder 14 can be reduced. This point will be described in detail with reference to FIG. 7. In FIG. 7, reference symbol A indicates the distance between the objective lens 10 and the imaging mirror 13, and reference symbol B indicates the imaging mirror 13 and the CCD camera. The distance between 15 CCD light receiving surfaces is shown. Between the objective lens 10 and the imaging mirror 13 is a diffusion system, and between the imaging mirror 13 and the CCD camera 15 is a condensing system. The light spread angle θ in the diffusion system is equal to the light collection angle θ in the light collection system. Therefore, the shorter the distance A is, the shorter the distance B is, so the length of the imaging tube 14 can be reduced.
[0030]
Further, the fluorescent microscope 1 is not only used with a rack and pinion mechanism to convert the manual operation of the operation dial 27 into the linear operation of the filter cassette 11 but also has a circular shape on the rotary shaft 26 of the operation dial 27 exposed to the outside. A seal member 28 is attached, and flanges 30 and 30 are provided at both ends of the circular seal member 28. With the flanges 30 and 30, an opening edge of a hole in the top surface plate 37 of the base 35 constituting a part of the light shielding member. Therefore, it is possible to reliably prevent light from entering through the hole while having a relatively simple configuration.
[0031]
Second embodiment (FIGS. 10 to 12)
10 to 12 show a second embodiment of the present invention, FIG. 10 corresponds to FIG. 1 of the first embodiment, FIG. 11 corresponds to FIG. 4 of the first embodiment, and FIG. This corresponds to FIG. 12 of the first embodiment, but the other parts of the second embodiment are substantially the same as those of FIGS.
[0032]
In the first embodiment, the display window 39 is provided in the case 38, and the characteristics of the filter cassette 11 currently set are displayed through the display window 39. For example, the display window 39 is omitted, and a number of, for example, transparent pockets or display frames 45 corresponding to the number of the filter cassettes 11 are provided at equal intervals on the peripheral surface of the operation dial 27. By attaching a display film or display paper describing the characteristics of the cassette 11 so as to be removable, the characteristics of the currently set filter cassette 11 are displayed.
[0033]
According to the second embodiment, information related to the currently set filter cassette 11 can be provided to the operator without the problem of light shielding.
[0034]
As described above, the fluorescence microscope 1 according to the embodiment is provided with the light shielding box 40 and covers all the elements except the operation dial 27 with the light shielding box 40. Since it can work and can be reduced in size compared to the prior art, it can be used on a desk used in daily work.
[0035]
The preferred embodiment of the present invention has been described above by taking the inverted fluorescent microscope as an example. For example, the objective lens 10, the filter cassette 11, the epi-illumination unit 8, and the imaging device 14 are arranged upright above the stage 2. Since it becomes a type | mold fluorescence microscope, an upright type | mold fluorescence microscope can be made by employ | adopting the structure substantially the same as the structure mentioned above.
[0036]
Moreover, it replaces with the dial 27 operated in order to select arbitrary filter cassettes 11, and the selection operation of the filter cassette 11 can be automated by installing an electric motor in relation to the rotating shaft 26.
[0037]
In addition, a plurality of filter cassettes 11 arranged in a straight line are linearly operated to set an arbitrary filter cassette 11. Instead of this, a plurality of filter cassettes 11 are arranged in an arc shape to form an arc trajectory. You may make it set arbitrary filter cassettes 11 by moving along.
[0038]
8 and 9 illustrate a revolver type filter cassette system. The illustrated revolver type filter cassette system has horizontal disks 43, 44 disposed at the upper end of the rotation shaft 26, and a plurality of filter cassettes 11 are disposed at equiangular intervals around the axis L <b> 3 of the horizontal disk 43. . More specifically, the four filter cassettes 11 are arranged at intervals of 90 ° around the axis L3, and by operating the manual dial 27, the four filter cassettes 11 are rotated to move any filter cassette 11. It can be set.
[0039]
In the revolver type filter cassette system illustrated in FIGS. 8 and 9, the selection operation of the filter cassette 11 can be automated by installing an electric motor in association with the rotating shaft 26 instead of the manual dial 27.
[0040]
Further, the fluorescent microscope 1 is covered with the light shielding box 40 that covers the entire surface. However, as disclosed in Patent Document 2, a light shielding member that at least partially shields light around the stage is provided so that it can be used outside the dark room. It may be.
[Brief description of the drawings]
FIG. 1 is a perspective view showing an internal structure of an inverted fluorescence microscope according to a first embodiment.
FIG. 2 is a plan view of a filter cassette, an epi-illumination unit, and an imaging unit extracted from the top.
FIG. 3 is a side view of a filter cassette and a driving mechanism for moving the filter cassette in a horizontal linear manner.
FIG. 4 is a perspective view of an inverted fluorescence microscope with a light shielding box removed.
FIG. 5 is a perspective view of a case housing a filter cassette and a rack and pinion mechanism.
FIG. 6 is a perspective view of an inverted fluorescence microscope in a state where a light shielding box is mounted.
FIG. 7 is a diagram for explaining the effect of shortening the length of the imaging cylinder.
FIG. 8 is a plan view of a revolver type filter cassette system.
FIG. 9 is a side view of a revolver type filter cassette system.
FIG. 10 is a perspective view showing an internal structure of an inverted fluorescence microscope according to a second embodiment.
FIG. 11 is a perspective view of the inverted fluorescence microscope of the second embodiment with the light shielding box removed.
FIG. 12 is a perspective view of a case containing a filter cassette and a rack and pinion mechanism related to the second embodiment.
[Explanation of symbols]
1 Fluorescence microscope 2 Stage 3 Sample 7 Mercury lamp (epi-illuminator)
8 Epi-illumination section 10 Objective lens 11 Filter cassette 12 Epi-illumination cylinder 13 Imaging mirror 14 Imaging cylinder 15 CCD camera 22 Slider 24 Spur gear (rack and pinion mechanism)
25 Circular gear (rack and pinion mechanism)
26 Rotating shaft 27 Operation dial 28 Circular seal member 37 Base top plate 40 constituting a part of the light shielding member Light shielding box L1 Axis line of incident illumination tube L2 Axis line of imaging tube

Claims (2)

A stage on which the sample is placed ;
A transmission illumination unit disposed above the stage and irradiating the sample on the stage with transmission illumination;
An objective lens disposed below and adjacent to the stage ;
A filter disposed below and in the vicinity of the objective lens ;
An epi-illumination tube disposed on a first horizontal axis perpendicular to the filter ;
An epi-illuminator attached to an end of the epi-illumination tube and for irradiating the sample on the stage with excitation light ;
An imaging mirror disposed below and in the vicinity of the filter;
An imaging cylinder arranged on a second horizontal axis arranged facing the imaging mirror;
An imaging device attached to an end of the imaging cylinder,
The epi-illumination tube and the imaging tube are arranged in a V shape having an acute angle when the first horizontal axis and the second horizontal axis are viewed from above ,
The fluorescent microscope, wherein the epi-illuminator and the imaging device are arranged at a height position where they interfere with each other when viewed from the side .   The fluorescence microscope according to claim 1, further comprising a light shielding member that shields light at least around the stage so that the fluorescence microscope can be used outside a dark room.

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