JP2007108224A - Total reflection microscope - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a total reflection microscope having improved operability in the case of changing an observation condition etc. <P>SOLUTION: The total reflection microscope has an illumination optical system for condensing illuminating light emitted from a lamp light source 1 on the pupil surface 12 of an objective lens 11 through an aperture diaphragm 4, and illuminates a sample 14 with the total reflection of the condensed illuminating light, wherein the aperture diaphragm 4 includes the total reflection illumination aperture diaphragm with a plurality of apertures 4a, and the total reflection microscope includes: a selecting means for selecting the aperture corresponding to the magnification of the objective lens 11 out of the plurality of apertures of the total reflection illumination aperture diaphragm; and an optical path shifting means 23 for positioning the part having the highest light quantity of the lamp light source in the aperture position of the selected aperture. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a total reflection microscope that performs total reflection illumination via an objective lens.

Conventionally, a total reflection microscope using a lamp light source (for example, a mercury lamp, a xenon lamp, etc.) is configured to project light from the light source onto a ring-shaped opening arranged in the illumination optical system. Is configured to be detachable from the optical axis of the illumination optical system. Also, in this type of total reflection microscope, several types of neutral density filters for adjusting the brightness of illumination light are provided in the microscope body, and the brightness of the observation light is adjusted according to the observation conditions (patent) Reference 1).
JP 2004-302421 A

  However, in a conventional total reflection microscope, the aperture stop is inserted from the optical axis in order to change the position or shape of the arc aperture stop for total reflection illumination depending on the magnification or NA of the objective lens used when observing the specimen. It is necessary to remove and readjust, and the operation is complicated.

  The present invention has been made in view of the above problems, and an object thereof is to provide a total reflection microscope having improved operability when changing observation conditions and the like.

  In order to solve the above-mentioned problems, the present invention has an illumination optical system that condenses illumination light from a lamp light source on the pupil plane of an objective lens through an aperture stop, and uses the collected illumination light as a sample. A total reflection microscope that performs total reflection illumination, wherein the aperture stop includes an aperture stop for total reflection illumination having a plurality of apertures, and the magnification of the objective lens from among the plurality of apertures of the aperture stop for total reflection illumination A total reflection microscope comprising: a selection unit that selects the aperture corresponding to the aperture; and an optical path shift unit that positions a portion of the lamp light source having the highest light amount at the aperture position of the selected aperture. I will provide a.

  Further, the total reflection microscope of the present invention has a variable aperture stop for epi-illumination, and is a neutral density filter that is disposed on the lamp light source side or the sample side of the variable aperture stop and can be inserted and removed integrally with the variable aperture stop. It is preferable to have a diaphragm exchanging means provided with

  In the total reflection microscope of the present invention, the stop replacement means moves the aperture stop for the total reflection illumination in a plane substantially perpendicular to the optical axis of the illumination optical system, so that it is on the pupil plane of the objective lens. It is preferable to have a position adjusting means for adjusting the condensing position.

  Further, in the total reflection microscope of the present invention, the diaphragm replacement means includes means for adjusting an aperture diameter of the variable aperture stop for epi-reflection illumination, and an optical axis of the illumination optical system with the center of the aperture of the variable aperture stop as the center. It is preferable to have means for adjusting to the above.

  In the total reflection microscope of the present invention, it is preferable that the diaphragm replacement unit is a slider that slides in a plane substantially perpendicular to the optical axis of the illumination optical system.

  In the total reflection microscope of the present invention, the optical path shifting means is made of parallel plane glass, and the inclination of the parallel plane glass with respect to the optical axis of the illumination optical system can be changed corresponding to the magnification of the objective lens. It is preferable.

  In the total reflection microscope of the present invention, the optical path shifting means can be switched between a first position corresponding to the first magnification of the objective lens and a second position corresponding to the second magnification. Is preferred.

  In the total reflection microscope of the present invention, the selection unit selects the aperture according to the magnification of the objective lens by rotating the aperture stop for total reflection illumination around the optical axis of the illumination optical system. It is preferable.

  ADVANTAGE OF THE INVENTION According to this invention, the total reflection microscope which improved the operativity at the time of changing observation conditions etc. can be provided.

  Hereinafter, an embodiment of the present invention will be described with reference to the drawings.

  FIG. 1 shows a schematic configuration diagram of a total reflection illumination state in the total reflection microscope according to the embodiment of the present invention, and FIG. 2 shows an epi-illumination state in the total reflection microscope according to the present embodiment.

  In FIG. 1, the light from the lamp light source 1 is condensed by the collector lens 2, the light amount is adjusted by a light-reducing filter set 50 for adjusting the light amount, and the total reflection is disposed on the slider 30 by the first light collecting lens 3. The light is condensed on the arcuate opening 4a of the aperture stop 4 for illumination. Between the first condenser lens 3 and the aperture stop 4 for total reflection illumination, a parallel plane glass 23 for shifting the optical path of the illumination light is disposed in the slider 30. The parallel plane glass 23 is disposed at a predetermined angle. By tilting, the optical path of the illumination light is shifted and condensed on the aperture 4a of the aperture stop 4 for total reflection illumination arranged so as to be shifted from the optical axis I of the illumination optical system.

  The illumination light condensed on the opening 4 a enters the collimating lens 5 and becomes parallel light, the illumination range is limited by the field stop 6, the traveling direction is changed by the bending mirror 7, and the second condensing lens 8 is applied. Incident. The illumination light that has passed through the second condenser lens 8 passes through the excitation filter 9 in the fluorescence filter cassette 22, the excitation wavelength is selected, is reflected by the dichroic mirror 10, and is reflected by the second condenser lens 8. 11 is condensed in the total reflection illumination range of the pupil plane 12 and emitted from the objective lens 11 to the specimen 14 as total reflection illumination light. In this way, the illumination optical system is configured.

  A petri dish 13 is placed on a stage (not shown), and a specimen 14 to be observed and a culture solution 15 for preventing drying of the specimen 14 are contained therein. The specimen 14 is obtained by, for example, staining cultured cell or cell tissue sections with a fluorescent dye. Further, a cover glass 16 is attached to the petri dish 13 on the objective lens 11 side, and a specimen 14 is attached thereon. A dedicated oil 17 is filled between the tip of the objective lens 11 and the cover glass 16.

  The evanescent wave is generated on the surface of the cover glass 16 on the sample 14 side by the total reflection illumination light, the sample 14 is irradiated with the fluorescence, and the fluorescence generated from the sample 14 by being excited by the evanescent wave is condensed by the objective lens 11, The light passes through the dichroic mirror 10 in the fluorescence filter cassette 22, passes through the emission filter 19 that transmits only fluorescence, is imaged on the imaging surface 21 by the imaging lens 20, and is disposed on the imaging surface 21. The image is picked up by an image pickup device such as a CCD shown in the drawing and is observed on a monitor (not shown). In this way, the total reflection microscope is configured. The fluorescence filter cassette 22 composed of the excitation filter 9, the dichroic mirror 10, and the emission filter 19 can be replaced depending on the type of fluorescence to be observed.

  In FIG. 1, the optical axis of the objective lens 11 and the optical axis from the lamp light source 1 to the folding mirror 7 are substantially parallel, but in actual arrangement, the optical axis of the objective lens 11 is bent from the lamp light source 1 with respect to the vertical. The optical axis up to the mirror 7 is substantially horizontal.

  In the present embodiment, in order to achieve the total reflection illumination state, the position of the opening 4a of the aperture stop 4 for total reflection illumination is within the total reflection possible region of the pupil plane 12 in a plane perpendicular to the optical axis I. The illumination light is arranged so as to be shifted from the optical axis I of the illumination optical system so that the illumination light is condensed. Further, the inclination of the parallel flat glass 23 is adjusted so that the maximum light amount portion of the lamp light source 1 is irradiated to the opening 4a, and the light emitting portion of the lamp light source 1 is focused on the opening 4a. Yes.

  In the total reflection microscope of the present invention, the slider 30 is configured to be detachable from the optical axis I in a plane substantially perpendicular to the optical axis I of the illumination optical system. Further, as will be described later, the slider 30 is provided with a circular variable aperture stop 25 used for epi-illumination and a neutral density filter 27 for adjusting the amount of light on the lamp light source 1 side of the variable aperture stop 25 so that it can be inserted and removed integrally. It has been. By sliding the slider 30 in a plane perpendicular to the optical axis I, one of the optical member made up of the aperture stop 4 for total reflection illumination and the parallel plane glass 23 and the optical member made up of the variable aperture stop 25 and the neutral density filter 27 is used. This optical member is inserted into the optical axis I of the illumination optical system, so that the aperture stop 4 or the variable aperture stop 25 can be replaced with respect to the optical axis I. When the aperture stop 4 or the variable aperture stop 25 is not necessary, the slider 30 can be removed from the optical axis I of the illumination optical system. It is also possible to insert a slider of the same type equipped with another filter or the like into the optical axis I of the illumination optical system.

  FIG. 2 shows a state in which the slider 30 is switched from the total reflection illumination state to the epi-illumination state. Switching from the total reflection illumination state to the epi-illumination illumination state is such that the center of the circular variable aperture stop 25 coincides with the optical axis I of the illumination optical system in a plane substantially perpendicular to the optical axis I of the illumination optical system. To move. At this time, the neutral density filter 27 configured to be insertable / removable is also inserted into the optical axis I of the illumination optical system at the same time so that a change in light quantity due to replacement of the aperture stop can be suppressed. Other configurations are the same as those in the total reflection illumination state of FIG.

  Next, the slider 30 used in this embodiment will be described. FIG. 3 shows a partial cross-sectional view of the slider 30 used in the present embodiment. 4 shows a partial cross-sectional view along the line AA in FIG. 3, and FIG. 5 shows a partial cross-sectional view along the line BB in FIG.

  3 to 5, the slider 30 is a variable aperture stop 25 for epi-illumination, an insertable / removable neutral density filter 27 (see FIG. 1), and an aperture stop for total reflection illumination having arcuate openings 4 a and 4 b. 4 and the parallel flat glass 23, and positioning click grooves 61 and 62 are provided on the outer side wall portion thereof. It is inserted by sliding on the optical axis I of the illumination optical system, and the position of each aperture stop is roughly set by positioning click grooves 61 and 62.

  In the case of FIG. 1, the aperture stop 4 for total reflection illumination is inserted into the optical axis I of the illumination optical system and positioned by the positioning click groove 61. In the case of FIG. 2, the variable aperture stop 25 for epi-illumination is inserted into the optical axis I of the illumination optical system and positioned by the positioning click groove 62.

  The variable aperture stop 25 for epi-illumination can be adjusted in the aperture diameter by pushing and pulling the aperture diameter adjusting lever 63, and the operating screws 64a and 64b are operated so as to be circular on the optical axis I of the illumination optical system. The center of the shape opening can be correctly aligned.

  The aperture stop 4 for total reflection illumination has an arc-shaped opening having a shape corresponding to the magnification of a plurality of objective lenses mounted on the microscope. In the present embodiment, as an example, a case where there are two types of arc-shaped opening 4a for a 60 × objective lens and arc-shaped opening 4b for a 100 × objective lens will be described. Needless to say, 4b is not limited to these two types. Two or more arc-shaped aperture stops may be formed.

  The aperture stop 4 for total reflection illumination is configured to be rotatable in a plane perpendicular to the optical axis I of the illumination optical system with respect to the holding component 65, and the user rotates to the position of the opening window 65 a formed in the holding component 65. The two types of openings 4a and 4b are selected and arranged by operating the operation screw 4c. Note that a click mechanism (not shown) may be provided to position the selected opening.

  The movement of the holding component 65 in the vertical direction on the paper surface of FIG. 4 is restricted by fitting with the holding component 66, and the movement of the optical axis direction of the illumination optical system is restricted by the housing of the slider 30 and the restriction plate 67. Therefore, it can be moved only in the longitudinal direction of the slider 30. Since the movement in the longitudinal direction is pressed in the direction of the operation screw 69 by the compression spring 68, the position of the holding component 65 in the longitudinal direction of the slider 30 can be adjusted by the operation of the operation screw 69. The position of the opening 4a in the longitudinal direction of the slider 30 can be adjusted.

  The holding component 66 is configured such that the movement of the illumination optical system in the optical axis direction is restricted by the housing of the slider 30 and the limiting plate 67 and can rotate only in the short direction of the slider 30 around the rotation shaft 70. It has become. The holding member 66 is pressed against the housing of the slider 30 by the compression spring 71 in the direction of one long side of the slider 30, and the force of the holding member 66 causes the operating screw 69 fitted to the holding member 66 to be pressed. Thus, the holding component 66 is held in a stopped state when the operation screw 72 screwed into the housing of the slider 30 receives a pressing force. The position of the arcuate opening 4a selected by moving the holding component 66 in the short direction of the slider 30 by operating the operation screw 72 can be adjusted in the short direction of FIG. The same applies when the arc-shaped opening 4b is selected.

  With the configuration described above, the position of the selected arcuate opening 4a or 4b in the plane perpendicular to the optical axis I of the illumination optical system can be adjusted in the longitudinal direction of the slider 30 in FIG. The position of the slider 30 in FIG. 4 can be adjusted in the short direction by operating the operation screw 72, and the light from the selected arcuate opening 4 a or 4 b is placed at a predetermined position on the pupil plane 12 of the objective lens 11. It can be condensed.

  Next, a description will be given of how to locate the place with the highest light quantity of the lamp light source 1 in the selected arcuate opening 4a (or 4b) when performing total reflection illumination. In the present embodiment, a parallel plane glass 23 for shifting the optical path by a predetermined amount from the optical axis I of the illumination optical system is disposed on the lamp light source 1 side of the aperture stop 4 for total reflection illumination of the slider 30. Yes.

  As shown in FIG. 5, the plane-parallel glass 23 is rotatably supported on the housing of the slider 30 by a rotation shaft 73 substantially perpendicular to the optical axis of the illumination optical system provided on the holding component 76. As shown in FIG. 3, the operation lever 74 can be rotated by the operation lever 74 attached to the holding component 76. The rotation amount is limited by the limit pins 75a and 75b, and the operation lever 74 is connected to the limit pins 75a and 75b. The portion with the highest light amount of the lamp light source 1 can be positioned at the opening position of the arc-shaped opening 4a or 4b corresponding to the objective lens 11 having a different magnification.

  For example, in the case of the objective lens 11 having a magnification of 60 times, the arc-shaped opening 4a is selected and positioned on the opening window 65a, and the operation lever 74 of the parallel plane glass 23 is positioned on the limit pin 75b side, whereby the lamp light source 1 is selected. The portion with the highest light quantity can be positioned in the arcuate opening 4a. Thereafter, the operation screws 69 and 72 shown in FIG. 4 are operated to collect the light from the lamp light source 1 at a predetermined position on the pupil plane 12 of the objective lens 11 so that the sample 14 can be totally reflected. When the objective lens 11 having a magnification of 100 times is replaced, the rotary operation screw 4c is operated to place the arcuate opening 4b in the opening window 65a, and the operation lever 74 of the parallel flat glass 23 is moved to the limit pin 75a side. By positioning the lamp light source 1, the portion with the highest light quantity of the lamp light source 1 can be positioned in the arcuate opening 4b. Thereafter, the operation screws 69 and 72 are operated to collect the light from the lamp light source 1 at a predetermined position on the pupil plane 12 of the objective lens 11 so that the sample 14 can be totally reflected.

  As described above, in the present embodiment, the parallel flat glass 23 is moved by the operation lever 74 while the arcuate opening 4a or 4b is switched to the position of the opening window 65a in accordance with the magnification of the objective lens used for specimen observation. By switching, the optical path of the illumination light is shifted by a predetermined amount, and the fine adjustment of the position of the arcuate opening 4a or 4b can be easily performed with the operation screws 69 and 72, so that the total reflection illumination state can be easily realized. Can do.

  Further, since the arcuate opening 4a or 4b can be moved in a plane perpendicular to the optical axis I of the illumination optical system by operating the operation screw 69, the incident angle of the illumination light is changed from the total reflection illumination state. It is possible to easily shift to the oblique illumination state and vice versa.

  Further, when the slider 30 is slid to switch from the total reflection illumination state to the epi-illumination state, the neutral density filter is detachably disposed between the epi-illumination variable aperture stop 25 and the first condenser lens 3. Since the amount of light is adjusted by 27, the amount of illumination light in the total reflection illumination state and the epi-illumination state can be made substantially the same. For this reason, there is no need to perform the troublesome operation of readjusting the neutral density filter set 30 provided in the microscope body even when switching from the total reflection illumination state to the epi-illumination state as in the past, and the illumination state Variation in the amount of light in the observation optical system due to switching can be suppressed. Therefore, when changing the observation conditions, the aperture stop can be easily replaced, the position is adjusted, and the amount of light can be easily adjusted, so that the operability of the total reflection microscope can be improved.

  In the above description, the case where the variable aperture stop and the arc-shaped aperture stop are exchanged with the slider has been described. However, the slider is not limited to the slider but may be a rotating turret or the like. Further, the tilt angle with respect to the optical axis of the parallel plane glass is not limited to the above-mentioned two points, and it may be configured to be fixed at an arbitrary angle between the maximum tilt angle and the tilt angle of zero degrees. With this configuration, it is possible to better match the maximum light amount of the lamp light source with the arc-shaped opening.

  The above-described embodiment is merely an example, and is not limited to the above-described configuration and shape, and can be appropriately modified and changed within the scope of the present invention.

FIG. 1 shows a schematic configuration diagram of a total reflection illumination state in a total reflection microscope according to an embodiment of the present invention; The epi-illumination state in this Embodiment is shown. It is a fragmentary sectional view of the slider which switches the aperture stop used for this Embodiment. FIG. 4 is a partial cross-sectional view taken along line AA in FIG. 3. FIG. 4 shows a partial cross-sectional view along the line BB in FIG. 3.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Arc light source 2 Collector lens 3 1st condensing lens 4 Aperture stop for total reflection illumination 5 Collimating lens 6 Field stop 7 Bending mirror 8 Second condensing lens 9 Excitation filter 10 Dichroic mirror 11 Objective lens 12 Pupil surface 13 Petri dish
14 Specimen 15 Culture Solution 16 Cover Glass 17 Oil 19 Emission Filter 20 Imaging Lens 21 Imaging Surface 22 Fluorescence Filter Cassette 23 Parallel Plane Glass 25 Variable Aperture Diaphragm for Epi-illumination 27 Dimming Filter 30 Slider 50 Dimming Filter Set 61 , 62 Click groove 63 Diaphragm diameter adjustment lever 64, 69, 72 Operation screw 65, 66, 76 Holding parts 67 Limit plate 68, 71 Compression spring 70, 73 Rotating shaft 74 Operation lever 75 Limit pin

Claims (8)

An illumination optical system for condensing illumination light from a lamp light source on a pupil plane of an objective lens through an aperture stop, and a total reflection microscope that performs total reflection illumination on the sample of the collected illumination light,
The aperture stop includes an aperture stop for total reflection illumination having a plurality of apertures;
Selecting means for selecting the aperture corresponding to the magnification of the objective lens from a plurality of apertures of the aperture stop for total reflection illumination;
An optical path shifting means for positioning the highest light quantity portion of the lamp light source at the opening position of the selected opening;
A total reflection microscope comprising: Has a variable aperture stop for epi-illumination,
2. The total reflection according to claim 1, further comprising: a diaphragm replacement unit provided with a neutral density filter that is disposed on the lamp light source side or the sample side of the variable aperture stop and that can be inserted and removed integrally with the variable aperture stop. microscope.   The aperture exchanging means is a position adjusting means for adjusting the condensing position on the pupil plane of the objective lens by moving the aperture stop for total reflection illumination in a plane substantially perpendicular to the optical axis of the illumination optical system. The total reflection microscope according to claim 1, wherein the total reflection microscope is provided.   The aperture exchanging means includes means for adjusting an aperture diameter of the variable aperture stop for the epi-reflection illumination, and means for aligning the center of the aperture of the variable aperture stop with the optical axis of the illumination optical system. The total reflection microscope according to any one of claims 1 to 3.   5. The total reflection microscope according to claim 1, wherein the aperture replacement unit is a slider that slides in a plane substantially perpendicular to the optical axis of the illumination optical system.   6. The optical path shifting means is made of parallel plane glass, and the inclination of the parallel plane glass with respect to the optical axis of the illumination optical system can be changed corresponding to the magnification of the objective lens. The total reflection microscope according to any one of the above.   7. The optical path shifting means can be switched between a first position corresponding to a first magnification of the objective lens and a second position corresponding to a second magnification. Total reflection microscope.   The selection means rotates the aperture stop for total reflection illumination around the optical axis of the illumination optical system, and selects the aperture according to the magnification of the objective lens. The total reflection microscope according to any one of the above.

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