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Iron staining (IS) fluid (chemical staining method)

A technology of iron staining and staining solution, which is applied in the biological field, can solve the problems of long time required, affecting the efficiency of pathological diagnosis, and insufficiently clear cell structure, and achieve the effects of reducing time consumption, shortening staining time, and clear cell structure

Active Publication Date: 2015-12-09
SHANGHAI SUNBIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Counterstaining reagents usually use basic fuchsin, sand yellow, nuclear fast red, etc. After counterstaining, the cell structure is not clear enough, and it is difficult to distinguish red blood cells from monocytes and granulocytes. It is necessary to judge the content of iron Longer time, thus affecting the efficiency of pathological diagnosis

Method used

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  • Iron staining (IS) fluid (chemical staining method)
  • Iron staining (IS) fluid (chemical staining method)
  • Iron staining (IS) fluid (chemical staining method)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: The iron staining kit for bone marrow cells of the present invention

[0038] Stationary solution: pure methanol solution;

[0039] Iron staining liquid I (potassium ferrocyanide solution): Weigh 2g potassium ferrocyanide, dissolve it in distilled water and dilute to 100mL with a concentration of 20mg / mL;

[0040] Iron staining liquid II (hydrochloric acid solution): Take 2 mL of concentrated hydrochloric acid, add 98 mL of distilled water, and mix well, with a concentration of 2%;

[0041] Nuclear fast red counterstain solution: weigh 0.2g nuclear fast red, 5g aluminum sulfate, 50mg thymol, 0.02g TritonX-100, add distilled water to 100mL, stir, boil for 5min to fully dissolve, and filter after cooling for use.

Embodiment 2

[0042] Example 2: The iron staining kit for bone marrow cells of the present invention

[0043] Fixing solution: take 10ml formalin (commercially available 37-40% formaldehyde aqueous solution), weigh 0.452g sodium dihydrogen phosphate (NaH 2 PO 4 ·2H 2 O), 1.64g disodium hydrogen phosphate (Na 2 HPO 4 ·12H 2 O), add 90 ml of distilled water, dissolve and mix thoroughly to obtain a 10% neutral formalin solution.

[0044] Iron staining liquid I (potassium ferrocyanide solution): weigh 5g potassium ferrocyanide, add distilled water to dissolve, dilute to 100ml, concentration 50mg / mL;

[0045] Iron staining liquid II (hydrochloric acid solution): Take 5ml of concentrated hydrochloric acid, add 95ml of distilled water, mix well, and the concentration is 5%;

[0046] Nuclear fast red counterstain solution: nuclear fast red counterstain solution: weigh 0.5g nuclear fast red, 5g aluminum sulfate, 50mg thymol, 0.005g TritonX-100, add 100ml of distilled water, stir, boil for 5min to fully disso...

Embodiment 3

[0047] Example 3: Iron staining method of bone marrow cells of the present invention (staining vat staining)

[0048] 1. Using the kit of Example 1;

[0049] 2. Method

[0050] Fixation: Take a bone marrow thick and thin smear (one end of the glass slide is rich in bone marrow particles, and one end is pushed into a thin slice), add 3-5 drops of methanol to the smear, cover the bone marrow membrane, fix for 10 minutes, wash with water, and wait for it to dry.

[0051] Iron staining: Mix the iron staining solution II and iron staining solution I in equal volumes to prepare the staining solution, put the bone marrow smear into the dyeing tank containing the staining solution, and adjust the microwave power to 500W microwave for 30s, wash with water, and wait for drying.

[0052] Counterstaining: nuclear fast red counterstaining for 2 minutes, wash with water, wait to dry, seal with neutral gum, and observe with oil microscope.

[0053] 3. Analysis of dyeing results:

[0054] The staining re...

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Abstract

The invention relates to the technical field of biology, and discloses an iron staining kit containing nucleus fast red counter staining fluid and a staining method thereof. The nucleus fast red counter staining fluid in the iron staining kit includes a decontamination agent TritonX-100. The ingredient composition of the nucleus fast red counter staining fluid is adjusted, the decontamination agent TritonX-100 is added, hence, the types of cells can be distinguished more easily in the iron staining process of bone marrow cells compared with conventional nucleus fast red counter staining fluid, the cell structure is clear and iron inside the cells can be determined more accurately. Meanwhile, in the staining method, a microwave mode is used for replacing a conventional water bath mode, and thus the overall staining time is greatly shortened.

Description

Technical field [0001] The invention relates to the field of biotechnology, in particular to an iron (IS) staining solution (chemical staining method), that is, an iron staining kit and a staining method. Background technique [0002] Normal human bone marrow contains a certain amount of iron, which is stored in the form of ferritin and hemosiderin, and used by blood cells to synthesize hemoglobin. This iron is called "extracellular iron". The iron particles that exist in red blood cells are called "intracellular iron". There are three main types of iron particles in red blood cells: immature red blood cells containing iron particles are called "iron granulocytes", and mature red blood cells containing iron particles are called "iron. "Granulocytes" are cells in which iron particles surround the nucleus of the young red blood cells in a circular distribution, called "ring-shaped iron young red blood cells". [0003] Iron chemical staining can locate intracellular and extracellular...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/30
Inventor 谢永华冯晓丽许付
Owner SHANGHAI SUNBIO TECH
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