JP2898241B2 - Optical microscope and sample dish - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an optical microscope for mounting a sample such as a cell and observing the sample not only vertically but also horizontally, and a sample dish for the same.

[0002]

2. Description of the Related Art An optical microscope generally comprises a combination of an objective lens and an eyepiece. A sample placed on a stage is illuminated with light from a light source, and an image enlarged by the objective is enlarged by an eyepiece. A minute portion of about 1 / 10,000 mm (10 ^-4 mm) can be identified. In particular, a method of observing an objective lens so as to be in close contact with a sample plate directly below the sample dish in order to increase the magnification is used.

[0003] When observing a sample, particularly lymphocytes contained in cells, with such an optical microscope, a sample dish which is generally called a chalet and can be filled with water or a solution is used to maintain the activity of the cells. Can be The sample dish is provided with a circular groove for water in the periphery, cells are placed in the center of the dish, and observation is performed by an optical microscope from above vertically or below the sample dish.

[0004]

However, when observing cells using the above-mentioned conventional sample dish, the microscope can only observe from above or below the sample dish. But,
When observing lymphocytes contained in cells, it is not possible to observe not only two-dimensionally but also from the horizontal side to specify the detailed shape and position. Nevertheless, data may not be obtained accurately. In particular, when cells that have floated in water and have a natural shape have drained water from the sample dish, the state changes greatly and becomes flat, and data from the horizontal side cannot be obtained more and more.

SUMMARY OF THE INVENTION The present invention has been made to solve such a problem, and a sample dish which allows an optical microscope to observe a sample not only from directly above or directly below but also from a horizontal side and an optical dish using the same. It is an object to obtain a microscope.

[0006]

According to the present invention, there is provided, as a means for solving the above problems, an irradiation light from an optical microscope for a dish container.
One side that accepts is open, the upper part of the gentle curved surface is provided in the center of the inside , a circumferential groove is provided around it, and a reflecting mirror with a cross section inclined outward at a predetermined angle along the groove peripheral surface is attached Thus, a sample dish for an optical microscope was formed.

[0007] In view of the above, the sample dish is composed of an observation section for irradiating light from a light source and enlarging and observing the sample with an objective lens and an eyepiece, and the sample dish for mounting the sample. The sample dish is installed on a rotating plate, and the rotating plate is provided on a movable table. The sample dish is moved and rotated, and any point of the reflection mirror is placed under the observation section, so that the horizontal side of the sample can be observed freely. The microscope was constructed.

The above-mentioned sample dish is mainly used for observation from above a vertical point and from a horizontal side surface, but it can also be used for cutting.

When the sample dish is used for observation, the sample dish is moved and rotated and set so that the observation portion comes to an arbitrary point on the reflection mirror of the sample dish. Light from a light source is reflected by a reflection mirror to irradiate the sample, and the external appearance such as the shape of the sample is observed by an observation unit.

This makes it easy to accurately observe a cell image in a state where a sample such as a cell containing a lymphocyte is floating on a culture solution surface and to observe a transient phenomenon such as cell fusion or division.

[0011]

Embodiments of the present invention will be described below with reference to the drawings. FIG. 1 is an overall schematic diagram of an optical microscope system using a sample dish according to an embodiment. The optical microscope 1 has an eyepiece and an objective lens like the conventional one, condenses light from a light source 4 with a condenser lens 5, refracts the optical path with a semi-transparent mirror 6, and passes through the objective lens 3. The sample X on the chalet 10 is illuminated, and the sample X is observed by the illumination.

The chalet 10 is obtained from an optical microscope of a dish container.
One side for receiving the irradiation light is open, and a cell layer 11 gently curled up in the center of the inside thereof and a circumferential groove 12 for water or a solution are provided around the cell layer 11, and a peripheral wall of the chalet is provided. Is provided at the upper end with a circumferential reflecting mirror 13 whose cross section is inclined outward by a predetermined angle (45 ° in the illustrated example). The chalet 10 is mounted on a rotary plate 14, and the rotary table 14 is provided on a movable cart 15.

A pinion 16 is engaged with the rotating plate 14, and a screw-type advance / retreat mechanism 18 is provided at an end of a carriage 15 movable by a roller 17. The pinion 16 is moved forward / backward by a motor 19a. The mechanism 18 is configured to be rotatable and movable by a motor 19b. The motors 19a and 19b are controlled by a control circuit 20.

FIG. 2 is a detailed plan view of the chalet 10.
As shown, the reflection mirror 13 is arranged in a ring shape along the outer periphery of the chalet. Tilt angle is 45 ° -60
°, and different angles (45 °, 60 °
°) is prepared in advance and can be replaced freely. In order to make the tilt angle variable, the reflecting mirror 13 is divided into a plurality of parts (for example, equal to 12), a circular hinge is provided at the lower end, and the circular band passed through the upper end is rotated by turning the knob of the screw adjusting mechanism of the terminal. The angle may be adjusted by expanding and contracting (not shown). Reference numeral 21 denotes an antireflection member.

The optical microscope system of the embodiment having the above configuration is used as follows.

When observing the cells of the sample X placed on the chalet 10 with this optical microscope system, the image of the sample from directly above is conventional and need not be described. The feature of this optical microscope system is that the sample X can be observed from the horizontal side.

When the sample X is placed on the horizontal side (the inclination angle of the reflecting mirror is 4
When observing from an oblique side when it is other than 5 °), see FIG.
As shown in FIG.
3 is set so that the optical microscope 1 is located right above.
In this case, in order to prevent the sample from becoming invisible due to the reflected light of the light from the light source, the antireflection member 21 is manually moved in advance so as to be located on the opposite side of the optical path of the reflection mirror 13.

When the side surface of the sample X reflected by the reflection mirror 13 is observed, the rotating plate 14 is then moved to the motor 19a.
Rotate slightly to observe the side of sample X from different angles. By repeating this operation and rotating the rotary plate 14 once, the side surface of the sample X can be observed from the entire circumference.

The optical microscope system according to this embodiment can be used to determine, for example, the shape of lymphocytes contained in cells, especially when they are taken out of a culture solution, or when cells are fused or divided. Suitable for observational study of shape change. The size of the lymphocytes is about 10 μm, and various state changes can be observed with the optical microscope system.

The above is a case where the appearance of the sample X such as a cell is observed.
0 is also used to cut cells or the like of the sample X by laser light. In this case, the sample X is irradiated with a laser beam for cutting from diagonally above while observing the sample X from above or from the horizontal side surface.

As a laser knife, for example, a wavelength of about 10 μm, such as a CO ₂ laser, is not suitable for cutting cells and the like, and for example, about 6 to 6.5 μm is desirable for cutting cells and the like. However, a laser generator having such a wavelength (outside medium-wave light) cannot be obtained with a semiconductor laser or a gas laser, but can be obtained with a free electron laser by appropriately setting parameters such as the magnetic field strength. Therefore,
By cutting cells with such a laser beam, a transient phenomenon such as cell fusion or division is caused, and this can be observed with the above-mentioned microscope system.

The laser beam may be directly applied to the sample such as a cell from obliquely above, or may be cut by applying the laser beam from the horizontal direction through the reflection mirror of the sample dish.

Although the reflection type optical microscope system has been described in the above embodiment, it goes without saying that the optical microscope itself can also be applied to a transmission type.

[0024]

As described in detail above, the sample dish of the first invention has a center at the center where the irradiation light is received from the open side, a circumferential groove at the periphery, and a cross-section outward along the circumferential surface of the groove. Because it was equipped with a reflective mirror provided at an angle,
An advantage is obtained that the sample can be irradiated and observed from the side by the reflection mirror, and the sample can be cut by reflecting the laser beam.

According to a second aspect of the present invention, in the optical microscope, the sample dish can be moved and rotated with respect to the observation section so that the observation section set on an arbitrary point of the reflection mirror can observe the horizontal side of the sample. Therefore, there is an advantage that a more detailed external shape of the sample can be obtained by observing not only an image from directly above the sample but also an image from the horizontal side.

[Brief description of the drawings]

FIG. 1 is an overall schematic diagram of an optical microscope system according to an embodiment.

FIG. 2 is a plan view and a partial cross-sectional view of a chalet.

FIG. 3 is an explanatory diagram of an operation.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 Optical microscope 2 Eyepiece 3 Objective lens 4 Light source 5 Condensing lens 6 Translucent mirror 10 Chalet 11 Cell layer 12 Circular groove 13 Reflection mirror 14 Rotating plate 15 Moving trolley 16 Pinion 17 Roller 18 Advance / retreat mechanism 19a, 19b Motor 20 Control circuit

Continuation of the front page (72) Inventor Masato Yasumoto 4547-44 Tsuda, Hirakata-shi Inside the Free Electron Laser Laboratory Co., Ltd. (56) References JP-A-63-98619 (JP, A) JP-A-1-312992 ( JP, A) JP-A-3-29909 (JP, U) JP-A-48-21621 (JP, Y1) (58) Fields investigated (Int. Cl. ⁶ , DB name) G02B 21/00-21/36 G01N 1/28 C12M 1/34

Claims (2)

(57) [Claims] 1. Receiving irradiation light from an optical microscope for a dish container
One side is open, the upper part of a gentle curved surface is provided in the center of the inside , a circumferential groove is provided around it, and a reflecting mirror whose cross section is inclined outward at a predetermined angle along the groove peripheral surface is attached. Sample dish for optical microscope. 2. The sample dish according to claim 1, further comprising: an observation unit configured to irradiate light from a light source to observe the sample by an objective lens and an eyepiece, and a sample dish according to claim 1 for mounting the sample. Is an optical microscope that is mounted on a rotating plate, the rotating plate is provided on a moving table, and a sample plate is moved and rotated to set an arbitrary point of a reflection mirror under an observation unit so that a horizontal side surface of the sample can be freely observed.