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A kind of Helicobacter pylori staining liquid and staining method thereof
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A technology of Helicobacter pylori and staining method, which is applied in the field of Helicobacter pylori staining solution and its staining, and can solve problems such as eye fatigue and easy missed diagnosis
Active Publication Date: 2021-12-31
SHANGHAI BAOSHAN DISTRICT RENHE HOSPITAL
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However, the size of the needle tip of the stained HP can only be observed under a high-magnification microscope, and the methylene blue staining method is relatively light. After a large number of cases are detected, the eyes of the diagnostician are prone to fatigue and misdiagnosis.
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[0038] 1 Materials and methods
[0039] 1.1 sample
[0040] Selected 3 cases of positive HP confirmed by gastroscopic biopsy in Department of Pathology, Renhe Hospital, Baoshan District, Shanghai in 2017.
[0041] 1.2 Main reagents and equipment
[0042] Methylene blue (methylene blue), sodium borate (sodium tetraborate decahydrate), boric acid: all purchased from Sinopharm Chemical Reagent Company); distilled water (preparation room of our hospital); formaldehyde solution; 75% ethanol; 95% ethanol; absolute ethanol ; n-butanol; xylene; embedding paraffin (68-60℃); automatic dehydrator (Leica SP300S); tissue embedding machine (Leica EG 1150H); rotary microtome (Leica 2235); Taiwan; medical oven; digital slide scanner (Jiangfeng KF-PRO-005).
[0043] 1.3 Method
[0044] 1.3.1 Sample processing
[0045] Get 1 case and prepare 2 parts of histological slices, divide the improved methylene blue method (embodiment 1) and B group without improved methylene blue method (comparati...
Embodiment 2
[0055] Example 2: Reagents for the modified methylene blue method: Add 1500 mg of sodium borate to 500 mL of distilled water, stir until completely dissolved, then add 3000 mg of methylene blue, stir and mix thoroughly, filter, put directly into a sealed dyeing jar protected from light, and store at room temperature .
Embodiment 3
[0056] Example 3: Reagent of the modified methylene blue method: add 4,500 mg of sodium borate to 500 mL of distilled water, stir until completely dissolved, then add 9,000 mg of methylene blue, stir and mix thoroughly, filter, put directly into a sealed dyeing jar protected from light, and store it at room temperature .
[0057] 1.3.3 Staining steps
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Abstract
A Helicobacter pylori staining solution and a staining method thereof, the staining solution is composed of the following raw materials: 6-18mg / mL methylene blue, 3-9mg / mL sodium borate, and the rest is distilled water, with the mass-volume ratio mg / mL as the unit. In the present invention, the sample tissue section is immersed in the above-mentioned Helicobacter pylori staining solution and dyed for <10s; the stained sample section is washed with running water, and then soaked in 95% ethanol, absolute ethanol, n-butanol, xylene, dehydrated, and dehydrated Afterwards, it was wet-sealed with neutral gum to obtain the stained slide sample; HP in the stained slide sample was observed under a microscope, and the diagnosis and interpretation of HP was performed according to the "China Chronic Gastritis Consensus (2017, Shanghai)". The present invention replaces boric acid with sodium borate to prepare methylene blue dye solution for staining Helicobacter pylori, significantly shortens the dyeing time, has the advantages of saving time and labor, deep bacterial color, easy microscopic examination and the like, and meets the diagnostic standard.
Description
technical field [0001] The invention belongs to the field of biological tissue staining, and in particular relates to a Helicobacter pylori staining solution and a staining method thereof. Background technique [0002] Helicobacter pylori (HP) infection is related to the occurrence and development of gastritis, gastric ulcer and gastric cancer. Therefore, the early detection and treatment of HP is of great significance to the prevention of gastric cancer. The methylene blue method is a commonly used staining method in clinical pathology. The existing methylene blue dye solution contains methylene blue (3,7-bis(dimethylamino)phenothiazine-5-onium chloride, also known as "methylene blue") and boric acid. However, the size of the needle tip of the stained HP can only be observed under a high-magnification microscope, and the methylene blue staining is relatively light. After testing a large number of cases, the diagnostician is prone to eye fatigue and misdiagnosis. Contents...
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