JP2007233066A - Specimen fixing method, fixing device, and stage device - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain a sharp image from a living body that exhibits dynamic behavior, especially, that moves in short cycles. <P>SOLUTION: There is provided a fixing method for a specimen A, such that the specimen A, placed on a stage 1, is pressed against the stage 1 by the tension of a sheet-like member 12 that is extended across the specimen A. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a specimen fixing method, a fixing apparatus, and a stage apparatus.

In recent years, in biological research, it has become possible to visualize ion concentration, membrane potential, etc. with a fluorescent probe using an optical microscope. So-called in-vivo observation for observing organs and the like has been performed. In in-vivo observation, the observation target has movements such as pulsation and respiratory movement, so that the observation position shifts and the focus is likely to be blurred.
As a method for eliminating such observation position deviation and out-of-focus blur, an apparatus is known in which an entire microscope including an imaging unit is made to follow the movement of an observation target (see, for example, Patent Document 1).
JP-A-7-222754

  However, the apparatus of Patent Document 1 has a problem that it cannot be operated at high speed because it is necessary to drive an entire microscope having an imaging means and having a large weight. For example, when observing the heart, the heart rate of the rat is about 350 times / minute and the heart rate of the mouse is about 620 times / minute, and it is extremely difficult for the device of Patent Document 1 to follow these heart rates. Have difficulty.

  The present invention has been made in view of the above-described circumstances, and an object of the present invention is to provide a microscope image capturing apparatus that can obtain a clear image from a living body that exhibits dynamic behavior and moves in a particularly short cycle.

In order to achieve the above object, the present invention provides the following means.
The present invention provides a specimen fixing method in which a specimen placed on a stage is pressed against the stage and fixed by the tension of a sheet-like member stretched over the specimen.
According to the present invention, the specimen placed on the stage is fixed in a state of being pressed against the stage by the tension of the sheet-like member. Since the sheet-like member has a large area and does not apply a large pressing force locally to the specimen, the influence on the specimen can be reduced, and the soundness of the specimen can be maintained. Moreover, the periodic pulsation of the whole specimen can be suppressed by fixing the specimen with a sheet-like member having a large area. As a result, it is possible to prevent the observation image from being blurred due to pulsation and obtain a clear image.

In the above invention, the sheet-like member may be in the form of a band that partially covers the specimen.
By doing in this way, an effective site | part can be suppressed partially in order to suppress a pulsation by a strip | belt-shaped sheet-like member.
In the above invention, the sheet-like member may cover the specimen as a whole.

In the above invention, the sheet-like member may be transparent.
By doing in this way, it becomes possible to observe the observation site | part of a sample through the sheet-like member which has fixed the sample. As a result, the observation site can be observed while being directly fixed by the sheet-like member, and blurring of the observation image can be effectively suppressed.

Moreover, in the said invention, it is good also as a through-hole provided in the position corresponding to the observation site | part of a sample in the sheet-like member spanned over the said sample.
By doing in this way, it becomes possible to expose the observation site | part of a sample from the through-hole of the sheet-like member which has fixed the sample, and to observe directly. Thereby, as a sheet-like member, it is not restricted to a transparent thing, The thing of arbitrary materials is employable.

The present invention also includes a frame having a size surrounding a specimen placed on a stage, a sheet-like member stretched inside the frame, and a pressing unit that presses the frame toward the stage. A specimen fixing device comprising:
According to the present invention, the sheet-like member stretched on the inside of the frame body is pressed from the specimen mounted on the stage, and the specimen is sandwiched between the stage and the sheet-like member, and the frame is pressed by the pressing means. By pressing the body toward the stage, it is possible to increase the tension of the sheet-like member and fix the specimen.

In the above-described invention, the frame body may include a pair of frame-shaped plates that sandwich the sheet-shaped member, and a close-contact unit that removably contacts the frame-shaped plates.
By doing in this way, a frame and a sheet-like member can be integrated by pinching | interposing a sheet-like member between a pair of frame-like plates, and making both frame-like plates contact | adhere by a contact means. Moreover, the sheet-like member can be separated from the frame body by releasing the close contact state of the frame plate by the contact means. Thereby, it becomes possible to replace | exchange a sheet-like member easily.

Moreover, in the said invention, the said close_contact | adherence means is good also as being comprised by making one side of the said frame-shaped plate into a magnet and making the other into a magnet or a magnetic material.
By doing in this way, a pair of frame-like plates can be stuck together using the magnetic attraction force of a magnet, and a sheet-like member and a frame can be integrated easily. Further, the sheet-like member can be easily separated from the frame body by pulling the frame-like plate away against the magnetic attractive force of the magnet.

In the above invention, the sheet-like member may be transparent.
Further, a through-hole may be provided in a position corresponding to the observation site of the specimen in the sheet-like member that is stretched over the specimen.

The present invention also provides a stage apparatus including a stage on which a specimen is placed and any one of the specimen fixing apparatuses described above.
According to the present invention, the specimen is clamped between the sheet-like member supported by the frame and the stage by pressing the frame toward the stage by the operation of the specimen fixing device, and the tension of the sheet-like member The sample can be fixed easily and effectively.

  According to the present invention, there is an effect that a clear image can be obtained from a living body that exhibits dynamic behavior and moves in a short cycle. Further, there is an effect that images can be obtained continuously and continuously from a living body moving with a large amplitude without losing the field of view.

Hereinafter, a method for fixing the specimen A according to the first embodiment of the present invention will be described with reference to FIG.
As shown in FIG. 1, the method for fixing the specimen A according to the present embodiment fixes the specimen A made of a laboratory animal such as a mouse placed on the stage 1 by the tension of the sheet-like member 2. . As the sheet-like member 2, it is preferable to use a transparent wrap material. After covering the specimen A, the sheet-like member 2 is fixed to the stage by an arbitrary fixing means such as an adhesive tape (not shown) in a state where a predetermined tension is applied.

  According to the fixing method of the specimen A according to the present embodiment, the specimen A is covered with a wide area by the sheet-like member 2 made of a wrap material, and the specimen A is entirely pressed by the tension applied to the sheet-like member 2, and the stage 1 Can be pressed against. Thereby, the pulsation of the sample A can be reliably suppressed without applying a strong pressing force to the sample A locally. Therefore, it is possible to observe the specimen A in a resting state while maintaining the soundness of the specimen A during observation without imposing an excessive burden on the specimen A.

  Further, by using the sheet-like member 2 made of a transparent wrap material, the objective lens 3 is brought close to the specimen A as shown in FIG. 1 while the sheet-like member 2 for suppressing the pulsation of the specimen A is interposed. The observation site B can be observed. As a result, it is possible to prevent the observation region B from being displaced due to dynamic behavior such as the pulsation of the specimen A, and to obtain a clear observation image in which blurring is prevented.

  In the fixing method of the specimen A according to the present embodiment, the case where the specimen A is entirely covered with the single sheet-like member 2 and fixed to the stage 1 has been described, but instead of this, FIG. As shown, the specimen A may be partially covered with two or more strip-shaped sheet-like members 2. Even in this case, since the pressing force is applied to the specimen A over a wide area, it is possible to prevent an excessive pressing force from being applied to the specimen A.

Moreover, by doing in this way, the observation site | part B of the sample A can be exposed, without covering with the sheet-like member 2, and a clearer observation image can be acquired.
Further, when the specimen A is fixed to the stage 1 so as to cover the specimen A with the same sheet-like member 2 as in FIG. 1, as shown in FIG. 3, the through-hole 4 is formed in the sheet-like member 2. By covering the specimen A so that the through hole 4 coincides with the observation site B, the observation site B is exposed while the pulsation of the specimen A is suppressed by the tension of the sheet-like member 2 and observation is performed. It becomes possible.

If the observation site B can be exposed in this manner, the material of the sheet-like member 2 is not limited to a transparent material, and the sheet-like member 2 of any material can be adopted, and the specimen It is possible to select one having flexibility and strength optimal for fixing A.
As shown in FIG. 4, when the pulsation cannot be suppressed only by the sheet-like member 2, it may be supplementarily suppressed by a stabilizer 5 extended from the outside of the stage 1. Further, two or more stabilizers 5 may be arranged, or one having two or more support columns 5a may be adopted.

Next, a specimen A fixing device 10 according to an embodiment of the present invention will be described below with reference to FIGS.
As shown in FIG. 5, the fixing device 10 according to this embodiment includes a frame 11 having an opening 11 a, a sheet-like member 12 stretched on the opening 11 a of the frame 11, and the frame 11 as a stage. 1 and pressing means 13 for pressing toward 1.

As shown in FIG. 7, the frame 11 includes a pair of frame-shaped plates 14 and a clip (contact means) 15 that keeps the frame-shaped plates 14 in a close contact state. Yes.
As shown in FIG. 5, each frame-shaped plate 14 has a substantially square outer shape, and has an opening 11 a larger than the specimen A placed on the stage 1 at the center. In addition, a plurality of through holes 16 are provided in the peripheral portion of each frame-like plate 14 at positions that coincide with each other when they are superimposed on each other.

  The clip 15 is constituted by a leaf spring member that sandwiches the overlapped frame-shaped plates 14 in the thickness direction, and the frame-shaped plates 14 are pressed against each other with a pressing force determined by the spring constant, and maintained in a close contact state. It is supposed to be. In the present embodiment, the clip 15 is disposed on each side of the substantially square frame-shaped plate 14.

The sheet-like member 12 is made of, for example, a transparent wrap material. The sheet-like member 12 is sandwiched between two frame-like plates 14 and pressed by a clip 15. Thereby, the sheet-like member 12 is being fixed to the frame-shaped plate 14 so that the opening part 11a may be plugged up.
The pressing means 13 is composed of a plurality of bolts 17 inserted into through holes 16 provided in the frame-shaped plate 14 and screw holes 18 provided in the stage 1 for fastening the bolts 17.

The operation of the specimen A fixing device 10 thus configured according to this embodiment will be described below.
In order to fix the specimen A on the stage 1 using the fixing device 10 according to the present embodiment, as shown in FIG. 5, a sheet attached to the frame body 11 from above the specimen A placed on the stage 1. The member 12 is lowered and brought into contact with the specimen A. Then, the bolt 17 that is passed through the through hole 16 of the frame-like plate 14 is fastened to the screw hole 18 of the stage 1.

By gradually tightening the bolts 17 into the screw holes 18, the frame 11 is pushed down toward the stage 1, so that the tension of the sheet-like member 12 increases and the pressing force applied to the specimen A increases.
As a result, as shown in FIG. 6, the specimen A is pressed by the sheet-like member 12 over a wide area, and by adjusting the tightening of the bolt 17, the tension applied to the sheet-like member 12 is adjusted, so The specimen A can be fixed on the stage 1 with pressure. Therefore, it is possible to reduce the burden on the specimen A and perform observation while maintaining the soundness of the specimen A.

  In addition, since the sheet-like member 12 is made of a transparent wrap material, the observation site B of the specimen A can be observed with the sheet-like member 12 interposed. And according to fixing device 10 concerning this embodiment, shift of observation part B by dynamic behaviors, such as pulsation of sample A, can be prevented, and a clear observation image which prevented blurring can be acquired.

  Further, in the present embodiment, the sheet-like member 12 is sandwiched between the pair of frame-like plates 14 and fixed by the clip 15 so that the sheet-like member 12 is fixed to the frame-like plate 14. In this case, or when the sheet-like member 12 is broken, the sheet-like member 12 can be easily replaced by simply removing the clip 15.

  In the specimen A fixing device 10 according to the present embodiment, the frame body is provided with a pair of two frame-shaped plates 14 and a clip 15 that closely contacts these frame-shaped plates 14. The frame body 11 may be constituted by a single frame-shaped plate 14 and the sheet-like member 12 may be bonded to the frame-shaped plate 14.

  In addition, the close-contact means for bringing the pair of frame-shaped plates 14 into close contact with the clip 15 is configured. Instead, one of the frame-shaped plates 14 is configured with a magnet and the other is configured with a magnet or a magnetic material. The contact means may be configured by By doing in this way, both frame-shaped plates 14 can be closely_contact | adhered with a magnetic attraction force, and the state which pinched | interposed the sheet-like member 12 with the contact | adhesion force can be maintained.

Moreover, you may provide the through-hole 4 similar to FIG. 3 in the sheet-like member 12 stretched by the opening part 11a. A plurality of belt-like sheet-like members 2 similar to those in FIG. 2 may be stretched in the opening 11a.
Moreover, although the bolt 17 and the screw hole 18 mutually fastened were employ | adopted as a press means, it replaces with this and may employ | adopt other arbitrary fixing means.

Further, in the fixing device 10 according to the present embodiment, the substantially square frame-shaped plate 14 is adopted, but instead of this, as shown in FIG. 8, a substantially circular frame-shaped plate 19 is adopted. Also good.
By doing in this way, the tension | tensile_strength which generate | occur | produces in the sheet-like member 12 by pressing the sheet-like member 12 against the sample A can be generated substantially uniformly over the entire circumference. Thereby, local concentration of tension can be prevented and the sheet-like member 12 can be prevented from being broken.

Next, a stage apparatus 20 according to an embodiment of the present invention will be described with reference to FIG.
As shown in FIG. 9, the stage device 20 according to the present embodiment includes a stage 21 on which the specimen A is placed, and a fixing device 22 that is provided on the stage 21 and fixes the specimen A.

  The fixing device 22 includes a frame body 23 provided on the stage 21 so as to be swingable about a horizontal axis C, a sheet-like member 24 stretched so as to close the opening 23a of the frame body 23, A pressing means 25 for fixing the frame body 23 in a state where the specimen A is pressed by the sheet-like member 24 is provided. Similar to FIG. 5, the pressing means 25 includes a screw hole 26 provided in the stage 21 and a bolt 27 fastened to the screw hole 26.

  According to the stage apparatus 20 according to the present embodiment, the specimen A is placed on the stage 21, and the frame body 23 on which the sheet-like member 24 is stretched is swung from the position indicated by the chain line in FIG. 9 to the position indicated by the solid line. The bolt 27 inserted into the through hole 28 provided in the frame 23 is fastened to the screw hole 26 of the stage 21 to generate tension on the sheet-like member 24 and fix the specimen A to the stage 21. Can do. In this way, the number of bolts 27 to be fastened is small, and the sample A on the stage 21 can be fixed more easily.

It is a perspective view explaining the fixing method of the sample which concerns on one Embodiment of this invention. It is a perspective view which shows the modification of the fixing method of the sample of FIG. It is a perspective view which shows the other modification of the fixing method of the sample of FIG. It is a perspective view which shows the further another modification of the fixing method of the sample of FIG. It is a perspective view which shows the fixing device of the sample which concerns on one Embodiment of this invention. It is a perspective view which shows the state which fixed the sample on the stage with the fixing device of the sample of FIG. FIG. 6 is an exploded perspective view of the specimen fixing device of FIG. 5. It is a modification of the sample fixing device of FIG. It is a longitudinal cross-sectional view which shows the stage apparatus which concerns on one Embodiment of this invention.

Explanation of symbols

A Specimen B Observation site 1,21 Stage 2,12 Sheet-like member 4 Through hole 10,22 Fixing device 11,23 Frame body 13,25 Pressing means 14,19 Frame-like plate 15 Clip (contact means)
20 Stage device

Claims (11)

  A method for fixing a specimen, wherein a specimen placed on a stage is pressed against the stage by a tension of a sheet-like member that is stretched over the specimen.   The specimen fixing method according to claim 1, wherein the sheet-like member has a strip shape that partially covers the specimen.   The specimen fixing method according to claim 1, wherein the sheet-like member entirely covers the specimen.   The specimen fixing method according to any one of claims 1 to 3, wherein the sheet-like member is transparent.   The specimen fixing method according to any one of claims 1 to 4, wherein a through-hole is provided in a position corresponding to an observation site of the specimen in the sheet-like member stretched over the specimen.   Specimen fixing apparatus comprising: a frame having a size surrounding a specimen placed on a stage; a sheet-like member stretched inside the frame; and a pressing unit that presses the frame toward the stage. .   The specimen fixing device according to claim 6, wherein the frame body includes a pair of frame-shaped plates that sandwich the sheet-shaped member, and a close-contact unit that removably contacts the frame-shaped plates.   The specimen fixing device according to claim 7, wherein the contact means is configured by using one of the frame-shaped plates as a magnet and the other as a magnet or a magnetic material.   The specimen fixing device according to any one of claims 6 to 8, wherein the sheet-like member is transparent.   The specimen fixing device according to any one of claims 6 to 9, wherein a through-hole is provided in a position corresponding to an observation site of the specimen in the sheet-like member stretched over the specimen.   A stage apparatus comprising: a stage on which a specimen is placed; and the specimen fixing apparatus according to any one of claims 6 to 10.

Images