JP3667599B2 - Tissue staining device - Google Patents

Description

[0001]
[Industrial application fields]
The present invention relates to an apparatus for staining biological tissue used for microscopic examination.
[0002]
[Prior art]
A biological tissue sample to be microscopically examined is prepared through a number of processes such as tissue fixation, embedding, slicing, and staining. In particular, the staining process requires a large number of reagents, and is a work that requires a large number of processes and takes time and labor. Specifically, the tissue slice S (FIG. 1) sandwiched between the slide glass 1 and the cover glass 2 is stained. For staining, the direction of the slide glass 1 is aligned and the staining housed in the staining tank 3 is stained. Immerse in reagent D and shake (Figure 2). A plurality of staining reagents D are used to stain the tissue section S in a predetermined color tone. Each time the staining reagent D is replaced, a separate staining tank 3 is used, and immersion and shaking are repeated. Although this operation is a simple operation, it is complicated and takes a long time.
[0003]
[Problems to be solved by the invention]
In consideration of the simplicity of work, an automatic dyeing device that simulates manual work has also been developed. However, since the mechanism for moving and shaking the slide glass 1 for each staining tank 3 is complicated, the conventional automatic staining apparatus is very expensive. Therefore, apart from an organization that processes a large amount of samples every day, it is impossible to introduce an expensive automatic staining apparatus from an economical viewpoint at a normal laboratory level.
Further, in the method in which the slide glass 1 is immersed in the staining reagent D accommodated in the staining tank 3, the staining reagent D is periodically replaced because staining may be insufficient due to deterioration of the reagent. . In particular, when precious tissue is stained, in order to obtain certainty even if the staining reagent D is considered to be relatively fresh, the entire amount is often replaced with a new reagent with knowledge of waste.
[0004]
[Means for Solving the Problems]
The present invention has been devised to solve such problems. By adopting a method in which a staining reagent flows intermittently on a slide glass, the structure is simplified and a large number of tissue samples are reproduced simultaneously. An object is to provide a tissue staining apparatus capable of staining with good quality.
[0005]
In the tissue staining apparatus of the present invention, a rotating shaft extends vertically upward from a drive motor disposed at the center of the pedestal, and a specimen set container is accommodated in a pan disposed above the pedestal. The container includes a bottom wall inclined downward in the radial direction and a side wall extending in the circumferential direction, and a reagent discharge portion is formed at a lower corner between the bottom wall and the side wall. A plurality of glass slides mounted with tissue sections are arranged radially around the rotation axis with the plate surface direction orthogonal to the bottom wall.
A dripping nozzle is directed to the upper corner of the slide glass located on the rotating shaft side. A reagent bottle containing a plurality of staining reagents can also be connected to the dropping nozzle via an outflow pipe having a check valve. The staining reagent dropped on the slide glass from the dropping nozzle flows down radially outward along the plate surface of the slide glass by the centrifugal force generated by the rotation of the container.
[0006]
Embodiment
In the tissue staining apparatus according to the present invention, for example, as shown in FIG. 3, a plurality of support legs 11 are erected on the upper surface of a base 10 at equal intervals in the circumferential direction. The support 11 supports the pan 12. The upper opening of the pan 12 is covered with a cover 13 as necessary.
A drive motor 14 is provided at the center of the pedestal 10, and a rotating shaft 15 of the drive motor 14 projects vertically upward. A container 16 having a bottom wall 16a inclined downward along the radial direction is fixed to the rotating shaft 15. A large number of slits or holes are formed in the lower corner between the bottom wall 16a and the side wall 16b of the container 16 to form a reagent discharge part 16c.
A large number of the slide glasses 1 to which the tissue sections S are attached are arranged radially around the rotation axis 15 in a state where the plate surface direction is orthogonal to the bottom wall 16a.
[0007]
Staining reagent supply system 20 for staining tissue sections S is provided with a plurality of kinds of staining reagent D _1, D ₂ ··· D _n reagent bottle 21 ₁ which houses each kind of, 21 ₂ · · 21 _n Yes. Each reagent bottle 21 ₁ , 21 ₂ ... 21 _n is connected to an outflow pipe 23 via a floating valve 22. The floating valve 22, the flow of the staining reagent D _1, D ₂ ··· D _n to the outlet pipe 23 attempts to flow back into the reagent bottle 21 _1, 21 ₂ ·· 21 _n is prevented. Each outflow pipe 23 is provided with a check valve 24, and merges into a staining reagent outflow pipe 25. At the lower end of the staining reagent outflow pipe 25, a dropping nozzle 26 that is accommodated in the container 16 and that faces the corner of the slide glass 1 is attached.
[0008]
The staining reagent D is dropped from the dropping nozzle 26 toward the upper corner of the slide glass 1 while rotating the container 16 with the rotational power from the drive motor 14. For example, open the reagent bottle 21 _first check valve 24 which contains a staining reagent D _1, by closing the other check valve 24, staining reagent D ₁ is dropped on the slide glass 1 from dropping nozzle 26.
[0009]
The dropped staining reagent D flows along the plate surface of the slide glass 1 because a radially outward force is applied by the centrifugal force F (FIG. 4) generated by the rotation of the container 16. As a result, a liquid film of the staining reagent D is formed on the slide glass 1. Since the tissue section S is usually 10 μm or less in thickness, it is sufficiently immersed in the liquid film of the staining reagent D and stained.
The staining reagent D that has come into contact with the tissue section S flows down toward the lower corner of the slide glass 1, and is collected from the pan 12 to the reagent collection bottle 27 through the reagent discharge part 16 c of the container 16. After the staining with the staining reagent D ₁ is completed, the check valve 24 for the reagent bottle 21 ₁ is closed, and the check valve for the reagent bottle 21 _{2 is} used to stain the tissue section S with another staining reagent (for example, D ₂ ). In the same manner, the staining reagent D ₂ is dropped onto the slide glass 1 from the dropping nozzle 26.
[0010]
Since the tissue section S is stained while the staining reagent D is allowed to flow intermittently on the slide glass 1 in this way, the staining is compared with the method in which the slide glass 1 is immersed in the conventional staining tank 3 (FIG. 2). The required amount of reagent D is greatly reduced. Also, even when stained tissue sections S at multiple staining reagent _{D 1, D 2 ··· D n} , without requiring replacement of the staining reagent _{D 1, D 2 ··· D n} , the check valve 24 staining reagent D ₁ that need by switching, D ₂ ··· D _n in tissue sections S is immersed. In addition, since the tissue section S is always in contact with the fresh staining reagent D, constant staining with good reproducibility can be obtained.
[0011]
【The invention's effect】
As described above, the tissue staining apparatus of the present invention employs a method in which a tissue section is immersed in a liquid film of a staining reagent that is generated by flowing a staining reagent onto a slide glass. A constant staining is achieved with good reproducibility. In addition, the consumption of staining reagents is greatly reduced, making it unnecessary to inspect and replace staining reagents that were required in the past, and even when multiple types of staining reagents are used, the entire process up to the end of staining is complete. Automation becomes easy.
[Brief description of the drawings]
FIG. 1 is a tissue section sandwiched between a slide glass and a cover glass. FIG. 2 is a schematic diagram illustrating a conventional tissue staining process. FIG. 3 is a tissue staining apparatus according to the present invention. Diagram explaining that reagent flows down radially outward by centrifugal force 【Explanation of symbols】
1: slide glass S: tissue section D, D ₁ , D ₂ ... D _n : staining reagent 10: pedestal 11: support leg 12: pan 13: cover 14: drive motor 15: rotating shaft 16: container 16a: bottom wall 16b: Side wall 16c: Reagent discharge part 20: Staining reagent supply system 21 ₁ , 21 ₂ ... 21 _n : Reagent bottle 22: Floating valve 23: Outflow pipe 24: Check valve 25: Staining reagent outflow pipe 26: Dropping nozzle 27: Reagent collection bottle

Claims (2)

A rotating shaft 15 extending vertically upward from a drive motor 14 disposed in the center of the pedestal 10 , and
Bread 12 arranged above the pedestal 10 , and
A bottom wall 16a fixed to the rotating shaft 15 and accommodated in the pan 12 , and having a bottom wall 16a inclined downward in the radial direction and a side wall 16b extending in the circumferential direction, between the bottom wall 16a and the side wall 16b A container 16 in which a reagent discharge part 16c is formed in the lower corner, and a plurality of glass slides 1 are arranged radially around the rotation axis 15 with the plate surface direction orthogonal to the bottom wall 16a ,
A drip nozzle 26 directed to the upper corner of the slide glass 1 located on the rotating shaft 15 side ,
Tissue characterized in that the staining reagent D dripped onto the slide glass 1 from the dripping nozzle 26 flows down radially outward along the plate surface of the slide glass 1 by centrifugal force generated by rotation of the container 16 Dyeing equipment. The tissue staining apparatus according to claim 1, wherein reagent bottles containing a plurality of staining reagents are each connected to a dropping nozzle through an outflow pipe having a check valve.

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