JPH02205773A - Hematoxylin staining solution - Google Patents

Abstract

PURPOSE:To obtain a hematoxylin staining solution excellent in a nucleus staining capacity, co-staining preventing properties and preservability and well usable especially in the staining of a thin section by preparing a composition consisting of hematoxylin of a specific wt. ratio, a mordant, an oxidizing agent, preservatives, an org. acid or an inorg. acid and the remainder of water. CONSTITUTION:At first, hematoxylin is used in an amount of 0.2-0.8% (on a wt. basis) and especially pref. in an amount of about 0.3% from the aspect of nucleus staining capacity. As a mordant, aluminum sulfate is pref. used and desirably used in an amount of 7-15%, especially, 9-12%. Further, an oxidizing agent is used in an amount of 0.04-0.06% and sodium iodide is well used most practically as the oxidizing agent. Furthermore, preservatives are used in an amount of 10-30%, pref., 15-25% in order to prevent the sedimentation of a suspended substance at the time of the preservation of a solution. As the preservatives or sedimentation preventing agent, glycerine can be used. Further, an org. or inorg. acid can be used in an amount of 0-20%, pref., 0.5-1.5% and the remainder is composed of water.

Description

DETAILED DESCRIPTION OF THE INVENTION [Technical Field] The present invention relates to a novel hematoxylin staining solution.

[Prior art]

Generally, when a specimen collected from a living body is subjected to a histological examination, the tissue cells are first fixed by immersing them in a suitable fixative for a certain period of time so as not to change them as much as possible. Then, it is placed in an embedding device and embedded in paraffin, and then cut into thin slices of a certain thickness using a microtome. The obtained section is stained by soaking it in a staining solution, and then placed on a piece of glass to make a specimen, which is then used for examination.

Hematoxylin staining solution is most often used as a staining solution when preparing this tissue specimen. When using this stain, the nucleus is generally stained blue and can be clearly distinguished from the cytoplasm and fibers.

This staining solution has the ability to stain nuclei darkly and clearly.
At the same time, it is required to have good co-staining prevention properties that prevent things other than the nucleus from being dyed at the same time, and also to have good preservability so that it can be stored and used for a long time without forming a precipitate. Furthermore, in the past, specimens were generally examined using sections with a thickness of 5 to 8 μm, but in recent years, with the aim of increasing the amount of information, examinations have begun to be made even thinner, approximately 2 to 3 μm. Well-dyed scales are also required.

Several types of hematoxylin staining solutions have been available for quite some time, although the basic composition has not changed. Their compositions are summarized in Table 1 below.

Each of these staining solutions has its own advantages and disadvantages; for example, Mayer's and Carazzi's solutions stain 4-5 μm sections well in a short time, but 3-μm sections require staining time of 30 minutes or more, and each solution is used gradually. This is no longer the case. In addition, Harris's solution produces a deep and vivid stain overall, but it has the disadvantage of using mercury, and also, along with double-strength Latzi's solution, precipitates form during storage, so this is also not practical. Also, Gill's solution is not strong enough for 3μm sections;
It is hardly used for m.

[Purpose and structure]

Thus, the present invention solves the above-mentioned difficulties and provides a hematoxylin staining solution that has excellent nuclear staining ability, anti-co-staining properties, and storage stability, and is particularly suitable for staining thin sections of 2 to 3 μm. The purpose is to

According to many experiments and studies by the present inventor, this purpose was achieved by using a weight ratio of 0.2 to 0.8% hematoxylin and 7% mordant.
~15%, oxidizing agent 0.04~0.06%, preservative 10~
It has been found that this can be achieved with a new hematoxylin staining solution consisting of 30% organic or inorganic acid, 0-2.0% organic acid or inorganic acid, and the balance water.

[Detailed description of the invention]

The present invention will be explained in more detail. In the present invention, % means weight %.

First, hematoxylin is used in an amount in the range of 0.2 to 0.8%, and in particular, from the viewpoint of nuclear staining ability, around 0.3% is preferable. Although ammonium ammonium and potassium ammonium are also used as mordants, they are poorly soluble, and in the present invention, readily soluble aluminum sulfate is preferably used. This mordant is 7
It is desirable to use ~15%, especially 9-12%, and most preferably to use a width of about 10.5%. Furthermore, in order to acidify the hematoxylin solution and dye it, an oxidizing agent of 0,0
Amounts of 4-0.06% are used. As the oxidizing agent, sodium iodine membrane is most often used practically, but hydrogen oxide (oxyflu) and potassium permanganate can also be used.

Furthermore, a preservative is used in an amount of 10 to 30%, preferably 15 to 25%, to prevent floating substances from settling during storage of the liquid. As the preservative or anti-settling agent, glycerin, chloral hydrate, ethylene glycol, etc. can be used. Furthermore, organic acids such as acetic acid and citric acid and inorganic acids such as hydrochloric acid are added in an amount of 0 to 2.0%, preferably 0.5 to 1.5%.
% can be used. If this acid or the above-mentioned mordant is used in a larger amount than above, co-dyeing can be effectively prevented, but the overall dyeability deteriorates, which is not preferable.

〔Example〕

Various hematoxylin staining solutions were prepared using various amounts of each of these compounds, and the preservability of the solutions was tested. The results are shown in Table 2 below.

As is clear from Table 2, the most preferred examples of the staining solution according to the present invention are as follows.

Hematoxylin 3.0g Water Approximately 700g Aluminum sulfate 105g Sodium iodate 0.6g Glycerin 200g Glacial acetic acid 5-15° Add water to 1,000cc This liquid has the following characteristics.

1) For 2-3 μm sections, sufficient nuclear staining can be achieved in about 10 minutes.

2) There is little co-staining and no sorting is required after nuclear staining.

8) It has good storage stability, with no floating matter occurring even after 20 weeks after production, and almost no change in dyeability.

These characteristics are that they are superior to conventionally widely adopted prescriptions in each of the items (■), 2), and 3). It will be like this.

Omotemiya Kara Harris 2x Kara Gil Present invention light Tsutsui nuclear staining ability 115415 Co-contamination prevention property 442 4 25 Preservability 44
1 3 25 [Effects of the Invention] As is clear from the above, when using the hematoxylin staining solution of the present invention, thin sections of 2 to 3 μm can be well stained in a short time, and the nucleus is well stained and co-staining is prevented. It is very practical because it has the effect of preventing precipitation and preventing precipitation even when stored for a long period of time.

Claims (1)

[Claims] Hematoxylin 0.2-0.8% by weight, mordant 7-
15%, oxidizing agent 0.04-0.06%, preservative 10-3
A hematoxylin staining solution consisting of 0% organic acid or inorganic acid 0-2.0% and residual water.