JP2009031688A - Coating composition for microscope slide glass, slide glass for microscope, preprocessing liquid for pam staining, and methenamine-silver stain solution for pam staining - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain a coating composition having a function for preventing or suppressing the collateral staining while keeping close contact with a pathologic tissue and cells, a slide glass for a microscope coated with the coating composition, a methenamine-silver stain solution for PAM staining having a function for preventing or suppressing the collateral staining at the PAM staining, and a preprocessing liquid for PAM staining. <P>SOLUTION: By using polyacrylamide as coating composition for microscope slide glass, hydrophilic property and close contact with the pathologic tissue and the cell on the surface of the slide glass is maintained, and the collateral stain is prevented or suppressed at the PAM staining. By dipping in a liquid containing the polyacrylamide before the ordinary PAM staining, or by mixing the polyacrylamide with the methenamine-silver stain solution for PAM staining, the collateral stain is prevented or suppressed at the time of PAM staining. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a coating composition for a microscope slide glass, a microscope slide glass using the same, a pretreatment liquid for staining, and a staining liquid. In particular, the present invention relates to a coating composition for a microscope slide glass effective for PAM staining, a microscope slide glass using the same, a pretreatment liquid for PAM staining, and a mesenamine silver staining liquid for PAM staining.

The microscope slide glass is required to have various functions such as affinity with a pathological tissue / cell, adhesion, and various stains, and the absence of a background (hereinafter referred to as co-staining) due to a staining method. Conventionally, slide glasses coated with aminosilane, poly-L-lysine, etc. have been widely used as slide glasses that require close contact with pathological tissues and cells. All of them are PAM (silver mesenamine periodate). ) At the time of dyeing, the mesenamine silver stain causes silver particles to adhere to the slide glass, resulting in co-staining, which hinders diagnosis. As a countermeasure, a method of adding gelatin to a mesenamine silver staining solution for PAM staining has been proposed (for example, see Non-Patent Document 1), but the effect is not sufficient. On the other hand, the uncoated slide glass does not co-stain at the time of PAM staining, but is insufficient in close contact with tissues and cells.
"Practical histopathology cytodiagnosis staining color picture book" Modern Publishing Co., Ltd. New edition March 15, 1999 Pages 31-35

  An object of the present invention is to provide a coating composition having a function of preventing or suppressing co-staining at the time of PAM staining while overcoming the above-mentioned drawbacks of the prior art and maintaining adhesion to pathological tissues / cells. The object is to provide a microscope slide glass coated on the surface, and a PEM staining mesenamine silver staining solution and a PAM staining pretreatment solution having a function of preventing or suppressing co-staining during PAM staining.

  In order to solve the above problems, a coating composition for a microscope slide glass containing polyacrylamide is provided. As a result, a coating composition having a function of preventing or suppressing co-staining during PAM staining while maintaining adhesiveness with pathological tissues / cells can be obtained.

  Further, if the microscope slide glass is coated with the coating composition, a microscope slide glass having a function of preventing or suppressing co-staining during PAM staining while maintaining adhesiveness with a pathological tissue / cell can be obtained.

  Moreover, the pretreatment solution for PAM staining containing polyacrylamide can prevent or suppress co-staining on the microscope slide glass during PAM staining.

  Moreover, the mesenamine silver staining liquid for PAM staining containing polyacrylamide can prevent or suppress co-staining on the microscope slide glass during PAM staining.

  As described above, polyacrylamide is used as a coating composition for microscope slide glass to prevent or inhibit co-staining during PAM staining while maintaining the hydrophilicity of the slide glass surface and adhesion to pathological tissues and cells. Is possible. Moreover, the co-dying at the time of PAM dyeing | staining can be prevented or suppressed by immersing in a polyacrylamide containing liquid before PAM dyeing | staining, or mixing polyacrylamide with the mesenamine silver dyeing liquid for PAM dyeing | staining.

  The coating composition in the present application refers to a coating composition containing poly-L-lysine, aminosilane, or both. Therefore, the coating composition as one embodiment of the present invention contains poly-L-lysine or aminosilane, or both of them, and polyacrylamide as a main component. Poly-L-lysine and aminosilane have the effect of improving the adhesion between the slide glass and the pathological tissue / cell. Polyacrylamide is effective in improving the hydrophilicity of the slide glass and preventing or suppressing co-staining during PAM staining. Poly-L-lysine is effective in improving the adhesion to pathological tissues and cells regardless of its molecular weight.

  Aminosilane is N-β (aminoethyl) γ-aminopropyltrimethoxysilane, N-β (aminoethyl) γ-aminopropyltriethoxysilane, γ-aminopropyltrimethoxysilane, γ-aminopropyltriethoxysilane, N- Phenyl-γ-aminopropyltrimethoxysilane and the like and hydrolysates thereof can be used.

  Polyacrylamide can be any of cationic, anionic and nonionic. The amount of polyacrylamide added is preferably 20% to 85% in terms of solid content of the coating composition. If it is less than 20%, the effect of preventing or suppressing co-staining during PAM staining is insufficient, and if it exceeds 85%, the pathological tissue / Adhesion with cells decreases.

Various additives can be added to the coating composition of the embodiment of the present invention. For example, metal oxides such as SiO ₂ , Al ₂ O ₃ , TiO _2, ZrO ₂ , and Sb ₂ O ₄ , various organoalkoxysilanes and hydrolysates thereof, alcohols such as ethyl alcohol and isopropyl alcohol, glycols, Examples thereof include water-soluble polymers such as polyvinyl alcohol, polyvinyl pyrrolidone, polyacrylic acid, and amino resins, and proteins such as saccharides and gelatin.

  The microscope slide glass as an embodiment in the present invention is obtained by coating the surface with the above coating composition, and the coating method is not limited.

  The mesenamine silver staining solution for polyacrylamide-containing PAM staining in the present invention is obtained by adding polyacrylamide to the mesenamine silver staining solution used for ordinary PAM staining. It is preferable to add 0.01 to 0.1 part. If it is less than 0.01 part, the effect of preventing or suppressing co-dying is insufficient, and if it exceeds 0.1 part, the mesenamine silver dyeing solution for PAM dyeing is emulsified and the dyeing becomes poor. The polyacrylamide-containing pretreatment solution for PAM staining as an embodiment of the present invention is a solution in which the test slide is immersed before the mesenamine silver staining in a normal PAM staining procedure, and contains polyacrylamide as a main component.

Examples of the present invention will be described below, but the present invention is not limited to this range.
(1) Examples and Comparative Examples for Coating Composition and Microscope Slide Glass in the Present Invention Example 1-1 Mix 100 parts of poly-L-lysine 0.05% aqueous solution and 2 parts of polyacrylamide 1% aqueous solution.
Example 1-2 100 parts of 0.05% poly-L-lysine aqueous solution and 25 parts of 1% polyacrylamide aqueous solution were mixed.
Example 1-3 A poly-L-lysine 0.05% aqueous solution 100 parts, a colloidal silica 1% aqueous dispersion 100 parts, and a polyacrylamide 1% aqueous solution 100 parts were mixed.
Example 1-4 100 parts of an aminosilane 1% aqueous solution and 100 parts of a polyacrylamide 1% aqueous solution were mixed.
Example 1-5 50 parts of an aminosilane 1% aqueous solution and 100 parts of a polyacrylamide 1% aqueous solution were mixed.
Comparative Example 1-1 Poly-L-lysine 0.05% aqueous solution Comparative Example 1-2 Poly-L-lysine 0.05% aqueous solution 100 parts and 1% gelatin aqueous solution 10 parts mixed Comparative Example 1-3 Aminosilane 1% aqueous solution Comparative Example 1-4 Mixing 100 parts of 1% aminosilane aqueous solution and 10 parts of gelatin

The evaluation method is shown below.
The coating compositions of each Example and Comparative Example were coated on an uncoated glass, dried and evaluated by the following method.
1. Hydrophilicity Visual observation of water repelling ○: No repelling Δ: Partially repelling ×: Repelling the entire surface Adhesiveness A pathological tissue section was placed on the slide glass, dried, autoclaved according to a conventional method, and stained with eosin to evaluate the adhesiveness between the slide glass and the pathological tissue.
○: No separation is observed Δ: Partial separation is observed ×: Separation is observed in a wide range PAM staining After the pathological tissue section was placed on the slide glass and dried, PAM staining was performed using the following mesenamine silver staining solution to evaluate the staining state and the presence or absence of co-staining.
Mesenamine silver staining solution 50 ml of 3% aqueous mesenamine solution, 5 ml of 5% aqueous silver nitrate solution, 5 ml of 5% aqueous sodium borate solution, and 40 ml of distilled water ○: No or little staining, and good staining Δ: Slightly co-staining Slightly unsatisfactory dyeing ×: Strong co-dyeing and poor dyeing

Table 1 shows the evaluation results obtained by the above evaluation method.

(2) Example of polyacrylamide-containing PEN-staining mesenamine silver staining solution and Comparative Example 2-1 In the present invention, 100 ml of the mesenamine silver staining solution was mixed with 2 ml of a 1% polyacrylamide aqueous solution. The slide glasses of Comparative Examples 1-1 and 1-3 in which pathological tissue sections were attached were immersed in this polyacrylamide-containing silver mesenamine staining liquid for PAM staining, and PAM staining was performed.
Example 2-2 100 ml of the above mesenamine silver staining solution was mixed with 4 ml of a 1% polyacrylamide aqueous solution and stained in the same manner.
Example 2-3 100 ml of the above-mentioned mesenamine silver staining solution was mixed with 8 ml of a 1% polyacrylamide aqueous solution and stained in the same manner.
Comparative Example 2-1 In Example 2-1, staining was performed without mixing a polyacrylamide aqueous solution.
Comparative Example 2-2 In Example 2-1, 2 ml of 1% gelatin aqueous solution was mixed and dyed instead of the polyacrylamide aqueous solution.

Example of polyacrylamide-containing pretreatment liquid for PAM staining in the present invention Example 3-1 Slide glasses of Comparative Example 1-1 and Comparative Example 1-3 with pathological tissue sections were added to a 0.1% polyacrylamide aqueous solution. After soaking for 5 minutes, it was stained with a usual PAM staining silver mesenamine stain without polyacrylamide.
The evaluation method was based on the evaluation method (1).

Table 2 shows the evaluation results obtained by the above evaluation method.

  In addition, when the microscope slide glass in the embodiment of the present invention is dyed using an automatic dyeing apparatus manufactured by Ventana, good staining results can be obtained. For polyacrylamide-free coated slides coated with aminosilane, etc., pre-treatment with skim milk, immunoblock (manufactured by Dainippon Sumitomo Pharma Co., Ltd.), etc. is required before staining with an automatic staining device manufactured by Ventana. When pretreatment is not performed, uneven dyeing or the like occurs, and good dyeing cannot be obtained. However, when the microscope slide glass according to the embodiment of the present invention is used, the pretreatment is not required and good staining is possible.

  As described above, polyacrylamide can be used as a coating composition for microscope slide glass to prevent or inhibit co-staining during PAM staining while maintaining hydrophilicity of the slide glass surface and adhesion to pathological tissues / cells. It becomes possible. In addition, by immersing in a polyacrylamide-containing solution before ordinary PAM staining, or by mixing polyacrylamide with a mesenamine silver staining solution for PAM staining, co-staining during PAM staining can be prevented or suppressed. Thus, the present invention is useful and can be widely applied to a microscope slide glass.

Claims (4)

  A coating composition for a microscope slide glass, comprising polyacrylamide.   2. A microscope slide glass obtained by coating the coating composition according to claim 1.   A pretreatment solution for PAM staining characterized by containing polyacrylamide.   A mesenamine silver staining solution for PAM staining characterized by containing polyacrylamide.