A kind of hematoxylin eosin staining solution and preparation method thereof

Abstract

The invention provides a hematoxylin eosin staining solution, comprising a hematoxylin staining solution and an eosin staining solution, wherein the hematoxylin staining solution comprises hematoxylin, an alumium salt, an oxidizing agent, benzalkonium chloride, an alcohol, a weak acid and water; the eosin staining solution comprises eosin, Biebrich scarlet, flame red B, water and ethanol. The invention also provides a preparation method of the hematoxylin eosin staining solution, and a dyeing method thereof. When the hematoxylin eosin staining solution provided by the invention is used for dyeing slices, the effects of clear background, distinct gradation, good dyeing effect, easy slice reading, not easy fading and convenience for long-term storage can be achieved.

Description

technical field [0001] The invention relates to a sample analysis pretreatment reagent, in particular to a hematoxylin and eosin staining solution, a preparation method thereof, and a staining method thereof. Background technique [0002] Hematoxylin and eosin staining, referred to as HE staining, is the most widely used technique in histology, embryology, and pathology. HE staining is to immerse tissue sections or cell smears in hematoxylin staining solution and eosin staining solution respectively, and after appropriate time and treatment, the tissues, cells and other components are stained with different shades of color, resulting in different refractive indices. It is convenient for observation and research under an optical microscope. [0003] Hematoxylin is a kind of basic dye, which is extracted from logan and is oxidized to generate hematin. It is used together with mordant to make the chromatin in the nucleus and the ribosome in the cytoplasm purple-blue. The mole...

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Problems solved by technology

However, the current hematoxylin staining solution has the following defects: 1) The oxidation process is easy to produce a metal film or the dyeing process is easy to cause dye precipitation, thereby contaminating the section, and in the production process, the above stains on the section are difficult to remove, resulting in staining. Inhomogeneity, or even unreadable slides, requires re-production; 2) False blue staining is prone to occur during the staining stage, and part of the cytoplasm is also stained blue, which affects the accuracy of subsequent observation results

However, all of the above-mentioned staining solutions using eosin as a staining agent have the following problems: 1) the composition of the dye is single, resulting in unclear layers when staining the cytoplasm and extracellular matrix; 2) due to the low staining intensity of eosin , the above-mentioned eosin dyeing solution needs to add acetic acid as a dyeing accelerator to enhance the dyeing strength, but the addition of the dyeing accelerator will lead to the weakening of the dyeing color level, which further aggravates the problem that the dyeing level is not clear enough; at the same time, adding the dyeing accelerator After the treatment of the slices, acidic substances need to be washed not only in the subsequent processing of the slices, but also when the acidic substances remain, it will easily fade during the preservation of the slices, and the obtained slices cannot be stored for a long time

[0012] Therefore, in the existing hematoxylin and eosin staining solution, the hematoxylin staining solution easily produces metal film or dye precipitation to make the dyeing uneven and prone to false blue staining, while the dyeing level of the eosin staining solution is unclear and the coloring effect is not good. In turn, the background of stained tissue sections or cell smears is blurred, making it difficult to read

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