JP2008139029A - Resin embedding auxiliary tool in preparing microscope sample, and resin embedding method using it - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a resin embedding auxiliary tool in preparing a microscope sample and a resin embedding method using it that can easily and certainly resin-embed a cultured cell cultured on a cover glass or the like. <P>SOLUTION: The auxiliary tool acted for resin embedding in preparing the microscope sample has an insertion hole where a capsule for resin embedding can be engaged and inserted into the center, has an adhesive layer on the bottom, and is made of synthetic resin such as EVA having flexibility. The resin embedding auxiliary tool is adhered to the cell surface on the cover glass on which the cell is cultured, the cell is arranged in the center hole of the resin embedding tool, the resin embedding auxiliary tool is engaged and inserted into an opening of the capsule for resin embedding into which the resin for embedding is previously input, and the cultured cell is embedded by the resin in the capsule for resin embedding. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a resin embedding auxiliary tool in the preparation of a microscope sample capable of easily and reliably embedding cultured cells cultured on a cover glass or the like, and a resin embedding method using the same.

  Conventionally, when observing cultured cells cultivated on cover glass, which is used as a research material in medicine and biology, or quasi-ultra-thin slices attached on slide glass, with an electron microscope, these observation objects are used. It was necessary to embed with resin in advance. In particular, the cells cultured on the cover glass are attached by extending the protrusions on the glass surface, so that they are removed by treatment with an enzyme such as trypsin for 5 to 10 minutes and then suspended by a centrifuge. It was necessary to collect in a lump form (pellet).

  However, when the resin embedding is performed by the above method, the cells are deformed into a spherical shape, which is different from the original form of the cells cultured on the glass. There was also a risk of mechanical damage of the cells due to centrifugation.

  Therefore, a method of embedding an embedding capsule into which an appropriate amount of epoxy resin or the like is poured by covering the cultured cell or quasi-ultra-thin section is also tried, but the cover glass or slide glass and the embedding capsule When the resin leaks from the gaps between them, or when the epoxy resin or the like is polymerized, the viscosity temporarily decreases, the embedding capsule moves on the glass surface, and the cultured cells or quasi-ultra thin sections are not embedded Has occurred. In particular, in the case of a low viscosity resin such as Lowicryl K4M (manufactured by Polysciences, Inc.) frequently used in histochemical studies using an electron microscope, it may leak due to a slight seismic intensity and thus cannot be used.

  Therefore, a plastic frame for preventing the antibody solution for immunohistochemical staining from flowing out has been proposed (see Patent Document 1).

Japanese Utility Model Publication No. 63-10454

  However, in the case of the plastic frame proposed in the above-mentioned Patent Document 1, an adhesive is attached to the back surface and detachably attached to the slide glass to prevent the antibody solution for staining from flowing out. At the time of observation, the glass surface is removed and the cover glass is used. However, the resin embedding is not considered by inserting the resin embedding capsule inside the frame, and the resin is polymerized. There is a problem in that the position is shifted due to a decrease in the viscosity of the resin when the resin is embedded, or when the embedded resin is removed from the glass surface after embedding, the resin can be removed together.

  In view of the above problems, the present inventors have made it possible to easily and easily cultivate cells cultured on a cover glass or the like without causing physiological changes or mechanical damage. It came to provide the resin embedding auxiliary tool in the microscope sample preparation which can embed resin reliably, and the resin embedding method using this.

  Therefore, the first feature of the auxiliary tool used for resin embedding in the preparation of a microscope sample of the present invention is that it has an insertion hole part into which a resin embedding capsule can be inserted at the center and an adhesive layer is formed at the bottom part. And

  In addition, the resin embedding auxiliary tool is characterized in that it is made of a synthetic resin having flexibility such as EVA.

  In addition, the resin embedding auxiliary tool is attached from the cell surface on the cover glass in which the cells are cultured, and the cells are placed in the central hole of the resin embedding tool, and the embedding resin is added in advance. The resin embedding method is a resin embedding method in which the resin embedding tool is inserted into the opening of the resin embedding capsule, and the cultured cells are embedded with the resin in the resin embedding capsule.

  A fourth characteristic is that the embedding resin is a low-viscosity resin.

  According to the resin embedding auxiliary tool in the preparation of the microscope sample according to the present invention, since the insertion hole part into which the resin embedding capsule can be inserted is formed at the center and the adhesive layer is formed at the bottom part, the cover glass or the like is used. It has an excellent effect that the resin embedding capsule can be securely fitted and inserted, and the resin embedding can be performed.

  Further, since the resin embedding auxiliary tool is composed of a synthetic resin having flexibility such as EVA, the resin embedding capsule in which cells are embedded after the resin is polymerized can be easily removed from the cover glass. Has an effect.

  Furthermore, the resin embedding auxiliary tool is attached from the cell surface on the cover glass in which the cells are cultured, and the cells are placed in the central hole of the resin embedding tool, and the embedding resin is added in advance. The resin embedding tool is inserted into the opening of the resin embedding capsule, and the cultured cells are embedded with the resin in the resin embedding capsule. It has an excellent effect that resin embedding can be surely performed without leakage.

  Hereinafter, the present invention will be described with reference to the drawings illustrating embodiments, but it is needless to say that the present invention is not limited to the embodiments. FIG. 1 is a plan view showing a resin embedding auxiliary tool in preparation of a microscope sample according to the present invention, FIG. 2 is a side view of FIG. 1, and FIG. 3 is an explanatory view of the use of the resin embedding auxiliary tool of the present invention.

  As shown in FIGS. 1 and 2, the resin embedding auxiliary tool 1 in the preparation of a microscope sample according to the present invention includes a ring body 2 having an insertion hole portion 3 in the center and an adhesive layer 4 on the bottom surface. Yes. The ring body 2 is made of a flexible EVA resin, and a material that strongly adheres to the glass surface is selected and used for the adhesive layer 4. The insertion hole 3 has a diameter into which an embedding capsule described later is fitted. In addition, the capsule for embedding used for a present Example was a product made from a polyethylene resin with the capsule projection part mentioned later.

  FIG. 3 shows an example of resin embedding using the resin embedding auxiliary tool 1 having the above-described configuration. The ring body 2 is placed with the cells 6 cultured on the slide glass 5 placed in the center. It is strongly bonded to the slide glass 5 by the adhesive layer 4. The resin embedding capsule 7 is inserted into the insertion hole 3, the opening 10 of the resin embedding capsule 7 is in contact with the surface of the slide glass 5, and the resin embedding capsule is placed on the upper surface of the ring body 2. 7 capsule projections 8 abut.

  When the resin 6 is embedded in the cells 6 with the above-described configuration, the embedding resin 9 previously filled in the embedding capsule 7 is embedded in the slide glass 5 and the ring body 2. The cell 6 can be reliably embedded without leakage of the embedding resin 9. In addition, even when the embedding resin 9 is polymerized, the ring body 2 and the slide glass 5 are strongly bonded by the adhesive layer 4 even when the viscosity is lowered, and the cells 6 can be securely embedded. . Furthermore, since the cells 6 are maintained in the state of being cultured on the slide glass 5 and are arranged at the center position of the ring body 2, they are suitable for observation with a microscope after resin embedding.

  Next, a resin resin embedding method using the resin embedding auxiliary tool 1 having the above configuration will be described with reference to FIG. First, a resin embedding capsule 7 is mounted so as to fit into the central insertion hole 3 of the EVA resin ring body 2, and the resin 9 used for embedding with the opening 10 facing upward, such as Lowicryl K4M (Polysciences, Inc.). Made)). Then, with the opening 10 of the resin embedding capsule 7 into which the resin 9 is injected facing upward, the cell 6 is positioned so that the cell 6 can be accommodated in the opening 10 as seen through the slide glass 5 to which the cell 6 has adhered. The slide glass 5 is pressed from above and strongly bonded with the adhesive layer 4. The slide glass 5 and the resin embedding capsule 7 thus bonded are turned upside down, and the cells 6 are embedded with the resin 9 in the resin embedding capsule 7. The resin 9 is polymerized by 360 nm ultraviolet light.

  When the embedding resin 9 is sufficiently polymerized, the resin embedding capsule 7 is extracted from the EVA resin ring body 2, the resin embedding capsule 7 is disassembled, and the embedded resin (not shown) is taken out. Sample for use.

  According to the resin embedding method using the resin embedding auxiliary tool in the preparation of the microscope sample of the present invention having the above configuration, the resin was cultured on a cover glass or the like without leaking even if the resin has a low viscosity. It is possible to embed cells with resin, and there is no adhesion failure due to low viscosity during polymerization of the resin. In addition, since the ring body can be attached after confirming the position of the cell on the cover glass or the like, the cell can be embedded at an optimal position as a microscope sample.

  The resin embedding auxiliary tool and the resin embedding method using the same in the preparation of a microscope sample according to the present invention can be utilized in a microscope sample preparation technique in the medical / biological field.

It is a top view of the resin embedding auxiliary tool in the microscope sample preparation which concerns on this invention. FIG. 3 is a side view of FIG. 2. It is use explanatory drawing of the resin embedding auxiliary tool in the microscope sample preparation which concerns on this invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Resin embedding auxiliary tool 2 Ring body 3 Insertion hole part 4 Adhesive layer 5 Slide glass 6 Cell 7 Resin embedding capsule 8 Capsule protrusion part 9 Embedding resin 10 Opening part

Claims (4)

  Auxiliary tool used for embedding resin in microscopic sample preparation, having an insertion hole into which a resin embedding capsule can be inserted in the center, and an adhesive layer formed on the bottom. tool.   The resin embedding auxiliary tool according to claim 1, wherein the resin embedding auxiliary tool is made of a synthetic resin having flexibility such as EVA.   The resin embedding auxiliary tool according to claim 1 or 2 is attached from the cell surface on a cover glass in which cells are cultured, and the cells are arranged in a central hole portion of the resin embedding tool, and are encapsulated in advance. A resin in microscopic sample preparation, wherein the resin embedding tool is inserted into an opening of a resin embedding capsule into which an embedding resin is charged, and the cultured cells are embedded with the resin in the resin embedding capsule. Embedding method.   4. The resin embedding method in preparing a microscope sample according to claim 3, wherein the embedding resin is a low viscosity resin.

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