JP4996973B2 - Stabilizer and biological observation apparatus - Google Patents

Description

  The present invention relates to a stabilizer and a living body observation apparatus.

  In recent years, in biological research, visualization of ion concentration, membrane potential, etc. using a fluorescent probe has been performed using an optical device. So-called in-vivo observation for observing organs and the like has been performed. In the in-vivo observation, the observation target has movements such as pulsation and respiratory movement, so that the observation position is easily shifted and the focus is blurred.

As a method of eliminating such observation position deviation and out-of-focus blur, an observation apparatus having means for restraining a sample exhibiting dynamic behavior is known (for example, see Patent Document 1).
This observation device includes a microscope observation device having an objective lens and a stabilizer that is brought into close contact with the sample, and the sample is pressed by the stabilizer to suppress the dynamic behavior of the sample.

JP 2005-338631 A

  However, in such a conventional observation apparatus, when observing an organ whose periphery such as the kidney is covered with other organs or fats, the outer shape of the organ contact surface of the stabilizer is larger than the outer shape of the objective lens. There is an inconvenience that other organs in the vicinity obstruct observation. That is, in order to bring the stabilizer into contact with the organ to be observed, it is necessary to push away other peripheral organs and tissues in advance, which is troublesome. In addition, if other organs cannot be pushed away, the stabilizer will press the organ to be observed together with other surrounding organs, etc., and sufficiently suppress the dynamic behavior of the organ to be observed. Inconveniently, a clear image cannot be obtained.

  The present invention has been made in view of the above-described circumstances, and in the observation of a living body exhibiting dynamic behavior, as represented by in-vivo observation, other organs exist around the sample. Another object of the present invention is to provide a stabilizer that can accurately hold a sample and suppress its behavior, and a living body observation apparatus that can easily obtain a clear image of the sample.

In order to achieve the above object, the present invention provides the following means.
The present invention relates to a stabilizer for being brought into close contact with the sample restrain movement of the sample, possess a contact surface which is brought into contact with the sample, and an outer peripheral surface tapering toward the contact surface, the outer peripheral surface A stabilizer capable of transmitting a pressing force to the contact surface via the contact surface and pressing the sample contacting the contact surface is provided.
According to the present invention, by bringing a contact surface into close contact with a sample to be observed, it is possible to suppress the dynamic behavior of the sample and observe a clear image without blurring.

  In this case, when the contact surface is brought closer to the sample, the outer peripheral surface tapered toward the contact surface is used to remove other organs or tissues existing around the sample to be observed. Since it moves while being pushed away by the outer peripheral surface, the contact surface can be brought close to and closely attached to the observation target. This eliminates the troublesome work of pushing away other organs in advance, and also prevents the inconvenience of pressing down other peripheral organs together, simplifying the dynamic behavior of the sample to be observed and It can be surely suppressed.

In the said invention, it is preferable that the said outer peripheral surface is formed in the taper shape.
In this way, other peripheral organs and the like can be displaced by sliding on the tapered outer peripheral surface, and the contact surface can be brought close to the observation target smoothly.

In addition, the present invention provides a living body observation apparatus including an optical device including an objective unit disposed in proximity to a sample, and the stabilizer that is brought into close contact with the sample at least around the observation range of the optical device.
According to the present invention, even if other organs exist around the observation range, the contact surface of the stabilizer can be easily brought close to the periphery of the observation range by the optical device and can be in close contact with the sample. The dynamic behavior is effectively suppressed, the fluctuation of the relative distance between the objective unit and the sample is reduced, and a clear image without blurring can be obtained by the optical device.

In the above invention, the objective unit has a tip portion formed in a tapered shape that gradually decreases in the observation direction, and the stabilizer includes a tapered inner surface that tapers toward the contact surface, and the taper The taper angle of the inner surface may be larger than the taper angle of the tip portion.
By doing in this way, it becomes possible to insert an objective unit easily inside a stabilizer, and an objective unit with a short working distance can be easily brought close to an observation object, and appropriate observation can be performed.

According to the stabilizer according to the present invention, in observation of a living body exhibiting dynamic behavior, as represented by in-vivo observation, the sample can be accurately obtained even if other organs exist around the sample. There is an effect that it can be suppressed and its behavior can be suppressed.
In addition, the living body observation apparatus according to the present invention has an effect that a sample can be accurately pressed by a stabilizer and a clear image of the sample can be easily obtained.

Hereinafter, a stabilizer 5 and a living body observation apparatus 1 according to an embodiment of the present invention will be described with reference to the drawings.
The stabilizer 5 according to the present embodiment is provided in the living body observation apparatus 1 as shown in FIG.

  As shown in FIG. 1, a living body observation apparatus 1 according to this embodiment mounts a sample A such as a mammalian cell such as an experimental small animal, a living tissue such as a muscle, or various organs such as a heart and a liver. A stage 2 to be placed, an optical device 4 provided on the stage 2, arranged opposite to the sample A on the stage 2, and provided with an objective unit 3 having a predetermined working distance, and arranged in the vicinity of the optical device 4 And a stabilizer 5 according to the present embodiment.

  The stage 2 includes an adjustment dial 6, and by operating the adjustment dial 6, the sample A can be moved in two horizontal directions, for example, the X and Y directions. Furthermore, the stage 2 is provided with a heater 2a. By heating the stage 2 with the heater 2a, the sample A placed on the stage 2 can be heated. Yes.

The optical device 4 is attached to a support column 8 extending in a vertical direction from a base 7 so as to be movable up and down by an elevating mechanism 9. By disposing the objective unit 3 vertically downward, the sample A on the stage 2 can be observed. Further, by operating the elevating mechanism 9, the objective unit 3 can be moved closer to and away from the sample A and the focus can be adjusted.
The objective unit 3 includes, at one end thereof, a distal end portion 3a formed in a tapered shape that gradually becomes thinner in the observation direction.

  The stabilizer 5 according to the present embodiment includes an arm 5a that is attached to a support column 10 that similarly extends in the vertical direction from the base 7 so as to be movable up and down by an elevating mechanism 11, and a tip portion 5b that is disposed at the tip of the arm 5a. I have.

  As shown in FIG. 2, the tip 5b is formed in a substantially frustoconical cylindrical shape with a small diameter on the sample A side, and has a frame 12 having an opening 12a at the tip, and the opening 12a of the frame 12 is closed. And a transparent disk-shaped glass plate 13 disposed at a position to be used. The front end surface disposed on the periphery of the opening 12a of the frame portion 12 and the surface of the glass plate 13 constitute a contact surface 5c that is brought into contact with the surface of the sample A.

The frame portion 12 has a certain axial length dimension (for example, about 3 mm or more), and the opening portion 12a is formed larger than the observation range by the objective unit 3. A tapered outer peripheral surface 12b and a tapered inner surface 12c taper toward the contact surface 5c are provided. As shown in FIG. 3, the tapered inner surface 12 c has a taper angle α that is larger than the taper angle θ of the distal end portion 3 a of the objective unit 3.
Thereby, when the contact surface 5 c is brought into close contact with the sample A, an observation range by the objective unit 3 is arranged in the opening 12 a of the frame portion 12.

Operations of the stabilizer 5 and the biological observation apparatus 1 according to the present embodiment configured as described above will be described below.
In order to observe the sample A of an organ such as an experimental small animal using the living body observation apparatus 1 according to the present embodiment, first, as shown in FIG. After the sample A such as the above is exposed, the elevating mechanism 11 is operated to lower the stabilizer 5, and the contact surface 5 c of the stabilizer 5 is brought into close contact with the surface of the sample A. In this state, the distal end portion 3a of the objective unit 3 is inserted inside the frame portion 12 of the stabilizer 5, the objective unit 3 of the optical device 4 is brought into close proximity to the glass plate 13, and focusing is performed. I do.

  Thereby, the dynamic behavior of the surface of the sample A pressed by the stabilizer 5, especially the dynamic behavior along the optical axis direction of the objective unit 3 is suppressed. Therefore, the observation range in the glass plate 13 in which the dynamic behavior is suppressed by the stabilizer 5 can be observed by the optical device 4, and a clear image without blur can be acquired.

  Further, according to the stabilizer 5 and the living body observation apparatus 1 according to the present embodiment, as shown in FIG. 3, when the periphery of the sample A to be observed is covered with another organ, fat D, or the like, When the tip 5b of the stabilizer 5 is brought closer to the sample A, the peripheral fat D and the like slide on the outer peripheral surface 12b with the movement of the tapered outer peripheral surface 12b. It will be pushed away. That is, the stabilizer 5 is smoothly moved toward the sample A while pushing away the fat D and the like around the sample A by the outer peripheral surface 12b of the frame portion 12 of the stabilizer 5, and the contact surface 5c of the stabilizer 5 is moved to the sample A. It can be easily adhered.

  As a result, the troublesome work of pushing away the fat D and the like in advance prior to the work of fixing the sample A by the stabilizer 5 becomes unnecessary. In addition, when the stabilizer 5 is brought into close contact with the sample A, the peripheral fat D and the like are not pressed together, so that the dynamic behavior of the sample A to be observed can be reliably suppressed. Furthermore, since the frame portion 12 of the stabilizer 5 has a certain height, it can be held so that peripheral fat D or the like covers the frame portion 12 and does not enter the observation range during observation. Therefore, peripheral fat D and the like can be prevented from obstructing observation, and a clear image without blurring can be obtained.

  Further, according to the living body observation apparatus 1 according to the present embodiment, since the taper angle α of the tapered inner surface 12c of the stabilizer 5 is larger than the taper angle θ of the distal end portion 3a of the objective unit 3, the distal end portion 3a of the objective unit 3 is moved. It can be easily inserted inside the frame portion 12. Therefore, observation can be performed with the objective unit 3 sufficiently close to the sample A without interfering with the stabilizer 5. Further, with such a configuration, the diameter of the tip of the frame portion 12 can be minimized. That is, since the contact surface 5c becomes small, even in the case of the sample A whose periphery is covered with fat D or the like as shown in FIG. 3, the dynamic behavior can be suppressed by reliably bringing the stabilizer 5 into close contact. .

  In the present embodiment, the material of the stabilizer 5 may be arbitrary, but from the viewpoint of reducing the invasiveness to the sample A such as an organ, the stabilizer 5 is made of a biocompatible material such as polysulfone. It is preferable to configure. Moreover, in order to improve visibility outside the observation range, the stabilizer 5 may be made of a transparent material.

Further, in the present embodiment, from the viewpoint of reducing the invasiveness to the sample A such as an organ, it is preferable that the edge portion of the distal end surface of the frame portion 12 is chamfered.
In order to avoid interference between the objective unit 3 and the frame portion 12, it is preferable that the thickness of the glass plate 13 be within the working distance of the objective unit 3.

  In the present embodiment, the glass plate 13 is provided in the opening 12a of the frame portion 12 of the stabilizer 5, and the tip surface of the frame portion 12 and the surface of the glass plate 13 are brought into close contact with the sample A as the contact surface 5c. However, instead of this, the glass plate 13 may not be provided, and only the front end surface of the frame portion 12 may be brought into close contact with the sample A as the contact surface 5c. Moreover, it may replace with the glass plate 13 and you may decide to provide another transparent member.

  In the present embodiment, the frame portion 12 is formed in a substantially truncated cone shape, but the shape of the frame portion 12 is not particularly limited, and any other shape is adopted. be able to. For example, it may be formed in a substantially triangular frustum shape as shown in FIG. 4 or a substantially square frustum shape as shown in FIG. Further, as shown in FIG. 6, the frame portion 12 may have a shape in which a substantially truncated cone portion and a substantially cylindrical portion are combined.

  Further, in the present embodiment, the outer peripheral surface 12b and the tapered inner surface 12c of the frame portion 12 of the stabilizer 5 are formed in a tapered shape, but instead of this, as shown in FIG. It may be formed so as to taper in steps.

Further, from the viewpoint of improving rigidity, as shown in FIG. 8, the outer peripheral surface 12b of the frame portion 12 may be formed to have a taper angle β larger than the taper angle α of the tapered inner surface 12c.
Further, as shown in FIG. 9, an uneven portion 12 d may be provided on the front end surface of the frame portion 12. With such a configuration, it is possible to increase the frictional force by causing the uneven portion 12d to bite into the surface of the sample A, and to suppress the behavior of the sample A in the XY direction.
As shown in FIG. 10, a member having a higher friction coefficient than the material of the stabilizer 5, for example, a ring-shaped sheet 12 e made of silicone may be provided on the front end surface of the frame portion 12.

  Further, as shown in FIG. 11, a suction groove 16 is provided on the front end surface of the frame portion 12, and a pipe connected to the suction groove 16 is configured by making the arm 5 a and the front end portion 5 b of the stabilizer 5 have a hollow structure. May be. In this case, as shown in FIG. 12, the living body observation apparatus 1 may include a suction pump 19 that sucks air through the tube 17 connected to the arm 5 a of the stabilizer 5.

  Then, the suction pump 19 is operated in a state where the front end surface of the frame portion 12 is in contact with the surface of the sample A so as to close the suction groove 16, so that the front end surface of the frame portion 12 is adsorbed to the sample A, Even when the displacement amount of A is large, the dynamic behavior of the sample A can be reliably suppressed.

In addition, as shown in FIG. 12, by providing a suction tank 18 between the stabilizer 5 and the suction pump, blood, mucus and the like can be prevented from entering the suction pump 19 even when an organ or the like is to be observed. Can do.
The shape and the number of the suction grooves 16 are not particularly limited. For example, as shown in FIG. 13, one substantially annular suction groove 16 may be provided, or a substantially circular suction groove may be provided. A plurality of them may be provided. In order to suppress excessive suction of the sample A such as an organ, a porous elastic member may be provided in the suction groove 16.

  In this case, a pressure sensor may be provided on the contact surface 5c of the stabilizer 5 and the suction pressure may be controlled based on the detected suction pressure. By doing in this way, the influence on sample A, such as an organ, by excessive aspiration can be prevented. Further, by providing a pressure sensor and detecting the suction pressure, the operator can confirm that excessive suction or incomplete suction has not occurred. In this case, the suction state may be displayed based on the suction pressure detected by the pressure sensor, or may be notified by sound or the like.

  In order to prevent excessive pressure, an excessive suction prevention mechanism may be provided. The excessive suction prevention mechanism is provided with a branch port in the tube 17, and prevents the suction pressure in the suction groove 16 from becoming excessive by opening and closing the tube. For example, the branch port may be opened and released to the atmosphere when the suction pressure of the suction groove 16 falls below a set lower limit pressure, and may be closed when the suction pressure rises.

  Further, in the present embodiment, the optical device 4 and the stabilizer 5 are separated from each other by being attached to the separate support columns 8 and 10 so as to be movable up and down. However, as shown in FIG. It may be attached. Alternatively, the stabilizer 5 may be attached to the arm 5 a that is fixed to the stage 2 and can be operated in the XYZ directions. By doing so, the stabilizer 5 can be operated together with the stage 2 on which the sample A is placed. Therefore, even if the sample A is moved by the operation of the stage 2 during observation, the same portion can always be suppressed by the stabilizer 5. .

  Further, as shown in FIG. 15, the stabilizer 5 may be configured integrally with the objective unit 3. In this case, when the tip 5b of the stabilizer 5 is brought into close contact with the sample A by adjusting the positional relationship between the objective unit 3 and the stabilizer 5 in advance according to the focal position of the objective unit 3, the objective unit 3 is simultaneously brought into contact with the sample A. Since the focal position of the lens is matched with the sample A, it is possible to save the trouble of performing the focal position adjustment every observation.

Moreover, in this embodiment, although the case where the heater 2a was provided in the stage 2 for mounting the sample A was described as an example, the present invention is not limited to this, and the heater 2a may not be provided. Alternatively, other heating means may be provided.
Moreover, you may decide to provide the stabilizer 5 so that the front-end | tip part 5b can be attached or detached with a screw | thread etc. with respect to the arm 5a. By changing the shape of the tip 5b in accordance with the type of the sample A, the sample A can be suppressed more appropriately and observation can be performed.

It is a whole lineblock diagram showing a living body observation device concerning one embodiment of the present invention. It is a longitudinal cross-sectional view which shows the structure of the front-end | tip part of the stabilizer which concerns on this embodiment with which the biological observation apparatus of FIG. 1 is equipped. It is a longitudinal cross-sectional view which shows the relationship between the sample in the biological observation apparatus of FIG. 1, and a stabilizer. It is a perspective view which shows the 1st modification of the stabilizer of FIG. It is a perspective view which shows the 2nd modification of the stabilizer of FIG. It is a longitudinal cross-sectional view which shows the 3rd modification of the stabilizer of FIG. It is a longitudinal cross-sectional view which shows the 4th modification of the stabilizer of FIG. It is a longitudinal cross-sectional view which shows the 5th modification of the stabilizer of FIG. It is a longitudinal cross-sectional view which shows the 6th modification of the stabilizer of FIG. It is a longitudinal cross-sectional view which shows the 7th modification of the stabilizer of FIG. It is a longitudinal cross-sectional view which shows the 8th modification of the stabilizer of FIG. It is a whole block diagram which shows the modification of a biological observation apparatus provided with the stabilizer of FIG. It is the front view which looked at the front-end | tip part of the stabilizer of FIG. 11 from the contact surface side. It is a whole block diagram which shows the modification of the biological observation apparatus of FIG. It is a whole block diagram which shows the other modification of the biological observation apparatus of FIG.

Explanation of symbols

A Sample B Observation range 1 Living body observation device 3 Objective unit 4 Optical device 5 Stabilizer 5c Contact surface 12b Outer peripheral surface 12c Tapered inner surface 13 Glass plate

Claims (4)

A stabilizer that is in close contact with the sample and restrains the movement of the sample,
Possess the contact surface is brought into contact with the sample, and an outer peripheral surface tapering toward the contact surface,
A stabilizer capable of transmitting a pressing force to the contact surface via the outer peripheral surface and pressing the sample that is in contact with the contact surface .   The stabilizer according to claim 1, wherein the outer peripheral surface is formed in a tapered shape. An optical device comprising an objective unit arranged close to the sample;
A living body observation apparatus provided with the stabilizer according to claim 1 or 2, which is brought into close contact with a sample at least around an observation range of the optical device. The objective unit has a tip formed in a tapered shape that gradually becomes thinner in the observation direction;
The stabilizer includes a tapered inner surface that tapers toward the contact surface;
The living body observation apparatus according to claim 3, wherein a taper angle of the tapered inner surface is larger than a taper angle of the tip portion.

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