JP2010008565A - Biological sample observation apparatus - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To observe biological samples while switching a low magnification objective lens and a high magnification objective lens without moving a visual field. <P>SOLUTION: The biological sample observation apparatus 1 includes: a stage 7 for mounting the biological samples A; a casing 10; a wide visual field objective lens 4 fixed on the casing 10 to condense light from the biological samples A and a high resolution objective lens 8 having higher magnification than the wide visual field objective lens 4; a casing moving means 11 for moving the casing 10; observation optical systems 10, 12 for observing light condensed by the wide visual field objective lens 4 and the high resolution objective lens 8; and an optical path switching means 6 arranged between the wide visual field objective lens 4, the high resolution objective lens 8 and observation optical systems 10, 12 and alternatively switching an optical path of the wide visual field objective lens 4 or the high resolution objective lens 8. The wide visual field objective lens 4 includes a flexible transmission member 4a transmitting images of the biological samples A on the edge, and the edge part 4b of the transmission member 4a is fixed on the stage 7. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to a biological sample observation apparatus.

  Conventionally, in a general optical microscope, when mounting multiple objective lenses with different magnifications to the revolver and changing the magnification (field of view) for observation, move the revolver to switch the objective lens to be placed in the observation optical path. Observe. The focal lengths of the objective lenses mounted on the revolver are almost the same, and even if the objective lenses are replaced, the sample to be observed and the objective lenses do not interfere with each other. In this way, it is possible to easily switch between macro and micro observations.

  Further, for example, in Japanese Patent Application Laid-Open No. 2005-234306 (Patent Document 1), a probe microscope for high-magnification observation and an auxiliary microscope for low-magnification observation are provided, and the two microscopes are moved along the same guide. There has been proposed a microscope observation apparatus capable of observing while switching between two microscopes without shifting the field of view by stopping the microscope at a predetermined position above.

JP 2005-234306 A

  However, in the case of the conventional general optical microscope and the microscope of Patent Document 1, in order to change the magnification and observe, it is necessary to first separate the low-magnification objective lens from the field of view and then change it to the high-magnification objective lens. For this reason, even if an attempt is made to observe a part specified by observation at a low magnification at a high magnification, the visual field may be lost. In particular, when observing a live mouse or the like, a desired observation site moves due to the pulsation or pulsation of the biological sample, and it is very difficult to observe while switching the magnification of the objective lens.

  The present invention has been made in view of the above-described circumstances, and an object thereof is to provide a biological sample observation apparatus capable of observing while switching between a low-magnification objective lens and a high-magnification objective lens without moving the visual field. Yes.

In order to achieve the above object, the present invention provides the following means.
The present invention relates to a stage on which a biological sample is placed, a casing, a wide-field objective lens that is fixed to the casing and collects light from the biological sample, and a high-resolution objective that has a higher magnification than the wide-field objective lens. A lens, a housing moving means for moving the housing, an observation optical system for observing light collected by the wide-field objective lens and the high-resolution objective lens, the wide-field objective lens, and the An optical path switching unit that is disposed between the high-resolution objective lens and the observation optical system and selectively switches the optical path of the wide-field objective lens or the high-resolution objective lens, the wide-field objective lens comprising: A biological sample observation apparatus is provided that includes a flexible transmission member that transmits an image of the biological sample at the tip thereof, and the tip of the transmission member is fixed to the stage.

  According to the present invention, a biological sample is placed on the stage, the focal point of the wide-field objective lens is focused on the biological sample, and a wide-field image is observed with the observation optical system to specify a portion to be observed in detail. When the high resolution objective lens fixed to the casing is moved to the specified observation site by the casing moving means and the optical path of the high resolution objective lens is selected by the optical path switching means, the field of view of the high resolution objective lens You can switch to and observe.

  In this case, the tip of the wide-field objective lens is made of a flexible transmission member, and the tip is fixed to the stage. The tip of the objective lens is not linked. That is, the high resolution objective lens can be moved within the field of view while observing the same field of view with the wide field objective lens. Thereby, the field of view of the high-resolution objective lens can be easily and accurately adjusted to a desired position.

  Further, the fields of the wide-field objective lens and the high-resolution objective lens can be switched by the optical path switching means without moving both objective lenses. As a result, the problem that the field of view is shifted or the observation site is lost can be solved by replacing the objective lens, and the wide-field objective lens can be easily fixed while fixing the field of view of the high-resolution objective lens at the desired observation site. The field of view of the high resolution objective lens can be switched.

  In addition, by observing a narrow range in detail with a high-resolution objective lens and then switching the field of view to a wide-field objective lens, which part of the biological sample was observed with the high-resolution objective lens, It can be easily confirmed. Further, this makes it possible to easily specify a detailed observation site, so that it is possible to observe the same site multiple times over time.

In the above invention, the optical path switching means may be a movable mirror.
In this way, the optical path can be switched with a simple structure, and at the same time, the optical path can be switched instantaneously because only the movable mirror is moved.

  According to the present invention, there is an effect that observation can be performed while switching between a low-magnification objective lens and a high-magnification objective lens without moving the visual field.

A biological sample observation apparatus 1 according to a first embodiment of the present invention will be described below with reference to FIG.
As shown in FIG. 1, the biological sample observation apparatus 1 according to this embodiment includes a laser scanning microscope 2, a monitor 3, a wide-field objective lens 4, and a support arm 5 that supports the wide-field objective lens 4. And an optical path switching means 6.

  The laser scanning microscope 2 includes a stage 7 on which a biological sample A is placed, a high-resolution objective lens 8, a laser light source 9, a scan unit (housing) 10, and a scan unit moving unit that moves the scan unit 10 ( (Case moving means) 11, a light detection circuit 12, and a computer 13.

A wide field objective lens 4 and a high resolution objective lens 8 are fixed to the scan unit 10.
The scan unit 10 includes an optical system for illuminating the biological sample A and observing an image therein. The laser light output from the laser light source 9 is reflected by the mirror 10a and the dichroic mirror 10b in the scan unit 10, and then is shaken by the galvano mirror 10d controlled by the galvano control unit 10c. The surface of the biological sample A is scanned through the image objective lens 8.

  Reflected light or fluorescence from the biological sample A is collected by the objective lenses 4 and 8, reflected by the galvanometer mirror 10 d, and part of the light passes through the dichroic mirror 10 b and is detected by the light detection circuit 12. And it is sent to the computer 13 as an electrical signal, and the observation image by each objective lens 4 and 8 is displayed on the monitor 3.

  The scan unit moving unit 11 controls the XY drive unit 11a that moves the scan unit 10 in the horizontal direction (XY direction), the Z drive unit 11b that moves the scan unit 10 in the focus direction (Z direction), and the drive units 11a and 11b. The XY control unit 11c and the Z control unit 11d are configured.

One end of the wide-field objective lens 4 is a fiber bundle 4 a, and the distal end 4 b is fixed to the stage 7 by a support arm 5.
The optical path switching means 6 is, for example, an optical path switching mirror 6a disposed at a position where the optical paths of the wide field objective lens 4 and the high resolution objective lens 8 in the scan unit 10 intersect, and is disposed outside the scan unit 10. The angle is controlled by the optical path switching control unit 6b.

  The optical path switching control unit 6b, the laser light source 9, the galvano control unit 10c, the photodetection circuit 12, the XY control unit 11c, and the Z control unit 11d are connected to the computer 13, respectively, and each operation is controlled by the computer 13. .

The operation of the biological sample observation apparatus 1 according to the present embodiment configured as described above will be described below.
In this embodiment, in order to observe the mouse B using the biological sample observation apparatus 1, the mouse B is placed on the stage 7 and the optical path switching mirror 6a is arranged at the position of the solid line in FIG. Then, laser light is output from the laser light source 9 and the surface of the mouse B is scanned, and a part to be observed in detail is determined while confirming the image of the wide-field objective lens 4 displayed on the monitor 3.

  Thereafter, while observing the wide-field image of the monitor 3, the scan unit 10 and the high-resolution objective lens 8 are moved by the scan unit moving means 11, and the tip of the high-resolution objective lens 8 is adjusted to the site to be observed. When the optical path switching mirror 6a is moved to the position indicated by the dotted line in FIG. 1 by the optical path switching control unit 6b, the field of view of the high resolution objective lens 8 is displayed on the monitor 3.

  In this case, the fiber bundle 4a provided at the tip of the wide-field objective lens 4 can be bent flexibly, and the tip 4b is fixed to the stage by the support arm 5. The same field of view can always be observed without being linked to the movement. Thereby, while observing the image of the wide-field objective lens 4, the high-resolution objective lens 8 can be moved within the field of view, and the high-resolution objective lens 8 can be easily and accurately adjusted to a desired observation site. There are advantages.

  In addition, since the optical path switching mirror 6a is moved by the optical path switching controller 6b to switch the visual fields of the wide-field objective lens 4 and the high-resolution objective lens 8, it is necessary to separate the visual fields of the objective lenses 4 and 8 from the desired parts. No, the positions of the objective lenses 4 and 8 can be held.

  Therefore, according to the present embodiment, the fields of the wide-field objective lens 4 and the high-resolution objective lens 8 can be easily switched while the fields of the objective lenses 4 and 8 are fixed in a desired range.

Further, according to the present embodiment, since the visual field can be switched instantaneously and accurately, detailed observation by the high-resolution objective lens 8 and confirmation of the position of the detailed observation site by the wide-field objective lens 4 can be smoothly performed. Can be repeated.
At the same time, since the observation site can be easily specified and aligned, for example, it is possible to observe the same site multiple times over time.

  In addition, since the observation images obtained by the high-resolution objective lens 8 and the wide-field objective lens 4 can be stored in one system, it is convenient for leaving the experimental results and also useful as data for the reproduction experiment. is there.

  Further, for example, when the abdomen of the mouse B is opened and the inside of the abdominal cavity is observed, a hole having a size large enough to accommodate the wide-field objective lens 4 and the high-resolution objective lens 8 is opened. Only 4 and 8 tips are inserted into the abdominal cavity. As a result, it becomes possible to observe the organs in the abdominal cavity, and it is not necessary to open the abdomen widely. Therefore, when observing a live mouse, damage to the mouse can be reduced.

A biological sample observation apparatus 1 according to a second embodiment of the present invention will be described below with reference to FIG.
The biological sample observation apparatus 1 in this embodiment uses a general upright microscope 14 instead of the laser scanning microscope 2 of the first embodiment described above, and uses a wide-field objective lens 4, a support arm 5, and an optical path. And a switching mirror 6a.

The upright microscope 14 includes a stage 7, a light source 15 that illuminates the biological sample A, a dichroic mirror 10 b, and a high-resolution objective lens 8.
The high-resolution objective lens 8 selects the field of view by moving the stage 7 and is moved in the Z direction by the focus handle 16 of the upright microscope 14.

The wide-field objective lens 4 is fixed to the upright microscope 14. Moreover, the fiber bundle 4a which can be moved flexibly similarly to 1st Embodiment is provided, and the front-end | tip part 4b is being fixed with respect to the stage 7 with the support arm 5. As shown in FIG.
The optical path switching mirror 6a is disposed at a position where the optical paths of the objective lenses 4 and 8 in the upright microscope 14 intersect.

  In order to observe the mouse B using the biological sample observation apparatus 1 according to the present embodiment configured as described above, the mouse B is illuminated by the light source 15 and the mouse B is observed through the eyepiece 17 by the wide-field objective lens 4. The detailed observation position is specified. The positions of the mouse B and the high-resolution objective lens 8 are adjusted by the stage 7 and the focus handle 16 so that the tip of the high-resolution objective lens 8 is arranged at a specific part, and the optical path switching mirror 6a is illustrated. When moved to the position of the dotted line 2, detailed observation with the high-resolution objective lens 8 can be performed with the eyepiece 17.

Thus, by using a general upright microscope 14 as a base, the mouse can be visually observed through the eyepiece 17. It is also possible to attach an image element such as a CCD camera 18 to the upright microscope 14 to acquire an observation image, or to construct a disk scan type confocal observation apparatus.
As the light source 15, a halogen lamp, a xenon lamp, a mercury lamp, or the like can be used.

1 is an overall configuration diagram showing a biological sample observation device according to a first embodiment of the present invention. It is a whole block diagram which shows the biological sample observation apparatus which concerns on the 2nd Embodiment of this invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Biological sample observation apparatus 2 Laser scanning microscope 3 Monitor 4 Wide-field objective lens 4a Fiber bundle (transmission member)
4b Tip of fiber bundle 5 Support arm 6 Optical path switching means 6a Optical path switching mirror (movable mirror)
6b Optical path switching control unit 7 Stage 8 High resolution objective lens 9 Laser light source 10 Scan unit (housing, observation optical system)
10a Mirror 10b Dichroic Mirror 10c Galvano Control Unit 10d Galvano Mirror 11 Scan Unit Moving Unit (Case Moving Unit)
11a XY drive unit 11b Z drive unit 11c XY control unit 11d Z control unit 12 Photodetection circuit (observation optical system)
13 Computer 14 Upright microscope (housing)
15 Light source 16 Focus handle (case moving means)
17 Eyepiece (observation optical system)
18 CCD camera (observation optical system)
A biological sample B mouse

Claims (2)

A stage on which a biological sample is placed;
A housing,
A wide-field objective lens that is fixed to the housing and collects light from the biological sample, and a high-resolution objective lens that has a higher magnification than the wide-field objective lens;
A housing moving means for moving the housing;
An observation optical system for observing light condensed by the wide-field objective lens and the high-resolution objective lens;
An optical path switching unit that is disposed between the wide-field objective lens and the high-resolution objective lens and the observation optical system and selectively switches an optical path of the wide-field objective lens or the high-resolution objective lens. ,
The biological sample observation apparatus, wherein the wide-field objective lens includes a flexible transmission member that transmits an image of the biological sample at a distal end thereof, and a distal end portion of the transmission member is fixed to the stage.   The biological sample observation apparatus according to claim 1, wherein the optical path switching unit is a movable mirror.

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