JPH11337830A - Microscopic objective lens and microscope - Google Patents

Abstract

PROBLEM TO BE SOLVED: To make a modulation contrast image and a phase difference contrast image rapidly and inexpensively observable by arranging a modulator and a phase ring on the pupil surface. SOLUTION: Vapor-deposited films 53a, 53b with different transmissivities as the modulator 53 and a circular phase ring 54 are provided on the surface of a parallel plane glass 52. The phase ring 54 is constituted of a phase film for phase shifting a beam by a 1/4 wavelength and an absorption film for decreasing an intensity of a direct beam and increasing contrast of an image. In such a manner, though the vapor-deposited films 53a, 53b and the phase ring 54 are provided on the parallel planar glass 52 of an objective lens, since the vapor-deposited films 53a, 53b and the phase ring 54 are a part of the parallel plane glass 52, they hardly are affected by that, and bright view observation is performed by an eyepiece.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a microscope objective lens and a microscope for observing a colorless and transparent specimen.

[0002]

2. Description of the Related Art Conventionally, a modulation contrast microscope has been known as a microscope for observing a colorless and transparent specimen (for example, cultured cells) (JP-A-51-128).
548 or JP-A-51-29149).

FIG. 7 is a diagram showing the principle of a modulation contrast microscope.

A modulation contrast microscope is
A slit plate 236 arranged in the pupil plane of the condenser lens 232, and the objective lens 25 conjugate to the slit plate 236.
And a modulator 253 that is disposed on the pupil plane 0 and has portions with different transmittances.

[0005] The modulator 253 is arranged such that portions having different transmittances coincide with the image of the slit 236a.

The light incident on the biological specimen 261 is refracted because the biological specimen 261 is curved (in FIG. 7, a truncated cone).

The light refracted by the biological specimen 261 passes through the objective lens 250 to form images at various positions. Each image has a high transmittance portion of the modulator 253 (for example, a transmittance of 100%).
%) A1, low transmittance portion (for example, transmittance 1% or less)
A2 and a portion where transmittance is intermediate (for example, transmittance 15%) A
3 respectively.

The images passing through the modulator 253 are simultaneously superimposed to give a relief-like contrast, so that an image 170 of a colorless and transparent biological specimen which is invisible can be visualized (modulation contrast image).

[0009] The modulation contrast microscope can also be used for transmitted illumination using a plastic or glass container.

[0010]

When observing a colorless and transparent biological specimen 261 such as a cultured cell, a modulation contrast image and a phase contrast image are conventionally used in many cases.

In order to observe a modulation contrast image, an objective lens 250 for modulation contrast is required, and in order to observe a phase difference contrast image, an objective lens for phase difference contrast is required.

Therefore, when the observation method is switched, the slit plate 236 must be replaced with an optical member having a ring stop, and the objective lens 250 must be replaced with an objective lens for phase difference contrast. I could not observe.

As the revolver, a revolver having four to five mounting holes is often used.
The objective lenses of ×, 10 ×, 20 ×, 40 ×, and 100 × are attached. However, in order to efficiently observe the modulation contrast image and the phase difference contrast image, 8 to 1 is required.
A revolver with 0 mounting holes is required,
Eight to ten objective lenses are required, resulting in high costs.

The present invention has been made in view of such circumstances, and an object of the present invention is to provide a microscope objective lens and a microscope capable of quickly and inexpensively observing a modulation contrast image and a phase difference contrast image. is there.

[0015]

According to a first aspect of the present invention, there is provided a microscope objective lens constituting an imaging optical system for forming an image of a sample placed on a stage. A modulator for observing a modulation contrast image and a phase ring for observing a phase difference contrast image are arranged on a pupil plane.

The phase contrast image can be observed by the modulator arranged on the pupil plane, and the phase contrast image can be observed by the phase ring arranged on the pupil plane.

According to a second aspect of the present invention, there is provided a microscope objective lens provided with a modulator for observing a modulation image of a specimen and a phase ring for observing a phase difference contrast image on a pupil plane, and the microscope objective lens. An illumination optical system for irradiating the sample with light from a light source, an illumination optical system having a first optical member having a modulation contrast slit, and a second aperture having a phase difference contrast ring stop. A movable holder for holding an optical member and selectively disposing the first optical member or the second optical member on a pupil plane of the illumination optical system.

The observation method can be switched simply by moving the movable holder and disposing the first optical member or the second optical member on the optical axis. Further, the modulation contrast image and the phase difference contrast image can be observed without switching the microscope objective lens.

[0019]

Embodiments of the present invention will be described below with reference to the drawings.

FIG. 1 is a side view of an inverted microscope according to an embodiment of the present invention.

The inverted microscope comprises a microscope body 10 and
Illumination column 20, condenser device (illumination optical system) 30
, A lens barrel 40, an objective lens (microscope objective lens) (imaging optical system) 50, and a stage 60.

The microscope body 10 has a base 11 and a base 1
1 and a support portion 12 formed at one end.

At the other end of the base 11, a lens barrel 40 is provided, and the lens barrel 40 is provided with an eyepiece 41 for enlarging an image generated by the objective lens 50 so that the image can be observed with the naked eye.

At the center of the base 11, an L-shaped vertical moving part 13 is provided.
The focusing handle 14 provided on the side surface of the “0” can move up and down.

A revolver 51 is mounted on the vertical moving unit 13, and a plurality of different types of objective lenses 5 are mounted on the revolver 51.
0 is attached.

On the pupil plane of the objective lens 50, a parallel plane glass 52 on which a modulator and a phase ring, which will be described later, are deposited.
Is arranged.

A stage 60 on which colorless and transparent living cells as a specimen 61 are set is provided above the objective lens 50, and both ends of the stage 60 are fixed to the support portions 12 and 15.

The lighting column 20 comprises a column 21 and this column 2
1 and a transmissive illumination unit 22 extending in the horizontal direction from the upper end.

A light source 23 is provided at the upper end of the column 21, and a condenser device 30 is provided at the transmission illumination section 22 via a mounting section 24.

The light emitted from the light source 23 is transmitted through the transmission illumination unit 22.
And the sample 61 is irradiated through the condenser device 30,
A modulation contrast image or a phase contrast image is formed by the objective lens 50, and the holder 16,
The sample is guided to the eyepiece 41 via the base 11 and the lens barrel 40, and the sample image is observed.

FIG. 2 is an enlarged sectional view of the microscope objective lens according to the present invention.

The objective lens 50 includes a cylindrical portion 55 and a lens 5.
6 and a parallel plane glass 52.

The cylindrical portion 55 holds a plurality of lenses 56 at predetermined intervals so that a predetermined magnification can be obtained. A magnification of about 1 to 100 can be obtained by combining a plurality of lenses 56.

The parallel flat glass 52 is arranged on the pupil plane of the objective lens 50.

FIG. 3 is an enlarged plan view of the parallel flat glass.

On the surface of the plane-parallel glass 52, as modulators 53, vapor-deposited films 53 a and 53 b having different transmittances and an annular phase ring 54 are provided.

The phase ring 54 is composed of layers of a phase film for shifting the phase of the light by 波長 wavelength and an absorbing film for directly reducing the intensity of the light and increasing the contrast of the image.

FIG. 4 is a sectional view of the capacitor device, and FIG. 5 is a plan view of the slider.

The condenser device 30 includes a cylindrical portion 31, a condenser lens 32, a slider 33, and a cylindrical frame.

The condenser lens 32 has an optical axis coinciding with the optical axis of the objective lens 50.

The slider 33 (movable holder) is disposed on the pupil plane of the condenser lens 32, and grasps the operation knob 33a or 33b and manually moves it linearly along the guide groove 35 in the vertical direction of FIG. .

The slider 33 has a first slit 36a having a rectangular slit 36a for modulation contrast.
Optical member 36 and phase difference contrast ring diaphragm 3
A second optical member 37 having 7a and an opening 38 for bright field observation are provided.

The first optical member 36 is formed of a light shielding material.
Light passes only through the slit 36a. The second optical member 37 is formed of a light shielding material, and light passes only through the ring stop 37a.

In the first optical member 36, a second polarizing plate 36b is disposed so as to cover almost half of the slit 36a.

The first optical member 36 is rotatably supported by the slider 33 and the slit 3 is
The direction of 6a can be fixed.

The cylindrical frame 34 is rotatably held in the opening 36c of the first optical member 36, and the first polarizing plate 34a is arranged at a position facing the opening 36c. Therefore, in the observation of the modulation contrast image, the modulation of the illumination light can be continuously changed.

V-grooves 33 c to 33 e engageable with a plunger 39 are provided on the side surface of the slider 33 for the first optical member 36.
It is formed at a position corresponding to the second optical member 37 and the opening 38.

The plunger 39 is urged toward the slider 33 by a spring (not shown).

A click mechanism is constituted by the V-grooves 33c to 33e and the plunger 39, and the first optical member 36, the second optical member 37, and the opening 38 are arranged on a reliable optical path by the click mechanism. .

Prior to observation, when the objective lens 50 is attached to the revolver 51 and the slider 33 is moved to dispose the first optical member 36 in the optical path of the condenser lens 32, the first optical member 36 is rotated. Accordingly, the position of the slit 36a is adjusted so that light is irradiated to a predetermined position (a range A indicated by a dotted line in FIG. 3) of the deposition films 53a and 53b.

Further, the objective lens 50 is attached to the revolver 51, and the slider 33 is moved to move the second optical member 37.
Is arranged in the optical path of the condenser lens 32, the position of the second optical member 37 is adjusted so that the image of the ring stop 37a is accommodated in the phase ring 54.

After the above adjustment, observation is performed.

When the slider 33 is moved to place the first optical member 36 in the optical path, the first polarizing plate 34
a, the light passing through the second polarizing plate 36a and the slit 36a enters the condenser lens 32 from an oblique direction with respect to the optical axis, and is converted into parallel light by the condenser lens 32 to the colorless and transparent biological specimen 61. Is irradiated from an oblique direction.

The light transmitted through the biological specimen 61 and incident on the objective lens 50 is applied to the range A of the deposition films 53a and 53b. At this time, a colorless and transparent image of the biological specimen is shaded by a difference in transmittance between the vapor-deposited films 53a and 53b, and a modulation contrast image of the specimen 61 is formed.

When the second optical member 37 is arranged in the optical path of the condenser lens 32 by moving the slider 33 by grasping the operation knob 33a or 33b, light passing through the ring stop 37a enters the condenser lens 32. Then, the sample 61 is irradiated.

The light is diffracted by the sample 61, and the diffracted light is out of phase by 1/4 wavelength with respect to the direct light. Since the light that has passed through the phase ring 54 has a １ ／ wavelength phase shift, for example, when the phase difference between the direct light and the diffracted light overlaps and the light intensities are strengthened, the sample 61 is a phase contrast image brighter than the background. Is formed.

When the slider 33 is moved by grasping the operation knob 33a or 33b to dispose the opening 38 in the optical path of the condenser lens 32, the light incident on the sample 61 through the opening 38 causes the sample 61 to pass through. The transmitted light is incident on the objective lens 50, and an image of the specimen 61 is formed by the objective lens 50.

At this time, the vapor deposition films 53a, 53b and the phase ring 54 are provided on the parallel flat glass 52 of the objective lens 50. The vapor deposition films 53a, 53b and the phase ring 54 are a part of the parallel flat glass 52. Therefore, the bright-field observation can be performed by the eyepiece lens 41 without being significantly affected.

Since the phase change cannot be detected with the naked eye, the transparent sample 61 cannot be seen in this bright-field observation.

FIG. 6 is a plan view of another slider, and FIG.
The same parts as those described above are denoted by the same reference numerals and description thereof will be omitted.

The slider 133 is provided with a third optical member 136 for observing a modulation contrast image in addition to the first optical member.

The third optical member 136 serves as the objective lens 50 for observing the modulation contrast image.
It is used when a special slit according to the magnification is required.

The third optical member 136 is also used for the slider 13
3 so as to be rotatable, and the direction of the slit 136a can be fixed by a clamp screw (not shown).

The V-grooves 33c-
33e, a V-shaped groove 133 engageable with the plunger 39.
c is formed at a position corresponding to the third optical member 136.

According to this embodiment, the following effects can be obtained.

By simply moving the slider 33 or the slider 133, the first optical member 36 and the second optical member 37 are moved.
Since it is not necessary to switch the objective lens 50, the modulation contrast image and the phase difference contrast image can be quickly observed, and the size of the revolver 51 is not increased.

Since it is not necessary to separately purchase the objective lens for the modulation contrast and the objective lens for the phase difference contrast, the economic burden on the observer is reduced.

When switching from the first optical member 36 to the second optical member 37, the first polarizing plate 34a is not disposed on the optical path, so that there is no shortage of light in observing the phase difference contrast image.

From the observation of the modulation contrast image using the first optical member 36, the third optical member 136
The first polarizing plate 134a and the second polarizing plate 1 even when switching to the observation of the modulation contrast image using
Since the relative position with respect to 36b does not change, there is no need to re-adjust the first polarizing plate 134a. In the above embodiment, the sliders 33 and 133 are used, but a turret may be used.

Further, in the above embodiment, the deposited films 53a,
Although 53b and the phase ring 54 are provided on the surface of the parallel flat glass 52, they may be provided on the surface of the lens 56 if it is a pupil plane.

[0071]

As described above, according to the microscope objective lens of the first aspect, the phase contrast image is arranged on the pupil plane by the modulator having the modulation contrast image arranged on the pupil plane. Can be observed respectively. As a result, the modulation contrast image and the phase contrast image can be observed with one microscope objective lens.
It is possible to perform quick observation, and it is not necessary to purchase an objective lens for the modulation contrast and an objective lens for the phase difference contrast as in the related art, thereby reducing the economic burden on the operator. .

According to the microscope of the second aspect, the observation method can be switched simply by moving the movable holder and disposing the first optical member or the second optical member on the optical axis. Quick observation can be performed. Also,
Since the microscope objective lens does not need to be switched, the modulation contrast image and the phase difference contrast image can be observed using a revolver of the same size having the same mounting hole as the conventional one.

[Brief description of the drawings]

FIG. 1 is a side view of an inverted microscope according to an embodiment of the present invention.

FIG. 2 is an enlarged sectional view of a microscope objective lens according to the present invention.

FIG. 3 is an enlarged plan view of a parallel flat glass.

FIG. 4 is a cross-sectional view of the capacitor device.

FIG. 5 is a plan view of a slider.

FIG. 6 is a plan view of another slider.

FIG. 7 is a diagram showing the principle of a modulation contrast microscope.

[Explanation of symbols]

 Reference Signs List 23 light source 30 condenser device (illumination optical system) 33 slider (movable member) 36 first optical member 36a slit 37 second optical member 37a ring diaphragm 50 objective lens (imaging optical system) 53 modulator 54 phase ring 60 stage 61 Specimen

Claims (2)

[Claims] A microscope for observing a modulation contrast image and a phase for observing a phase difference contrast image in a microscope objective lens constituting an imaging optical system for forming an image of a sample placed on a stage. A microscope objective lens, wherein a ring is arranged in a pupil plane. 2. A microscope objective lens provided with a modulator for observing a modulation image of a specimen and a phase ring for observing a phase difference contrast image on a pupil plane, and light which coincides with an optical axis of the microscope objective lens. An illumination optical system having an axis and illuminating the sample with light from a light source, and a first having a modulation contrast slit
A movable holder that holds the first optical member and the second optical member having a phase difference contrast ring stop, and selectively arranges the first optical member or the second optical member on a pupil plane of the illumination optical system. A microscope comprising: