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Leukocyte classification kit and classification method thereof

A white blood cell classification and white blood cell technology, applied in biological testing, material stimulation analysis, material inspection products, etc., can solve the problems of low quantum yield, increased detection cost, easy photobleaching, etc., achieve low reagent consumption and reduce detection cost Effect

Inactive Publication Date: 2013-12-04
嘉善加斯戴克医疗器械有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] U.S. Patent 6004816 discloses a method for five classifications of white blood cells, which is to label nucleic acid substances in white blood cells with cyanine fluorescent dyes under the action of a hemolytic agent, and then measure side scattered light and fluorescence intensity signals to classify and count white blood cell subpopulations , but this type of fluorescent dye is easy to photobleach and has a low quantum yield, which increases the amount of each detection and increases the cost of detection

Method used

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  • Leukocyte classification kit and classification method thereof
  • Leukocyte classification kit and classification method thereof
  • Leukocyte classification kit and classification method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1 water-soluble CdTe quantum dot-cyanine dye probe

[0047] (1) Preparation of glutathione-stabilized water-soluble CdTe quantum dots

[0048] a. Preparation of NaHTe solution

[0049] Install a 50mL clean three-neck flask on a magnetic heating stirrer, add 0.09g Te powder and 0.06g NaBH4 into it under nitrogen protection, keep the temperature at 75°C, and inject 3mL of boiled and cooled double distilled water. React for 10 minutes, until the black powder disappears, the solution turns into purple-red NaHTe solution, which is marked as A solution.

[0050] b. Preparation of glutathione-stabilized water-soluble quantum dots

[0051] Under nitrogen protection conditions, add 100mL 1.0mmol / L Cd to a clean 200mL three-neck flask 2+ solution and 0.1mmol / L glutathione (GSH) solution, adjust the pH to 10 with 1mol / L NaOH solution, then add 300uL of solution A, heat and reflux for 100min, and prepare glutathione-stabilized CdTe (CdTe-GSH) quantum dots.

[0052...

Embodiment 2

[0056] Embodiment 2 prepares leukocyte classification reagent (A)

[0057] Prepare a white blood cell differential reagent containing the following substances for testing.

[0058]

[0059] Add 10uL of EDTA.2K anticoagulated blood to 1mL of white blood cell classification assay reagent, then add 3uL of 0.2ppm water-soluble CdTe quantum dot-cyanine dye probe to prepare a test sample, and inhale the appropriately modified flow cytometry analysis The instrument uses the side fluorescence with a measurement angle of 90 degrees to measure the fluorescence intensity information of the cells, and uses the side scattered light with a measurement angle of 90 degrees to measure the side scattered light intensity information of the cells. The results are as follows figure 2 As shown, white blood cells are divided into five types: lymphocytes, monocytes, neutrophils, basophils and eosinophils.

Embodiment 3

[0060] Embodiment 3 prepares leukocyte classification reagent (B)

[0061] Prepare a white blood cell differential reagent containing the following substances for testing.

[0062]

[0063]

[0064] Add 10uL of EDTA.2K anticoagulated blood to 1mL of white blood cell classification reagent, then add 3uL of 0.3ppm probe water-soluble ZnTe quantum dot-cyanine dye probe to prepare the test sample, and inhale the properly modified flow The cell analyzer uses side fluorescence at a measuring angle of 90 degrees to measure the fluorescence intensity information of cells, and uses side scattered light at a measuring angle of 90 degrees to measure the side scattered light intensity information of cells. The results are as follows image 3 As shown, white blood cells are divided into five types: lymphocytes, monocytes, neutrophils, basophils and eosinophils.

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Abstract

The invention discloses a leukocyte classification and a classification method thereof, and the kit is composed of a water soluble quantum dots-cyanine dye probe, and an erythrocyte and platelet hemolytic agent, wherein the quantum dots-cyanine dye probe is composed of a stabilizing agent, which takes a polypeptides material as the quantum dots, and Cy5 cyanine dye covalent joint molecules whose emission spectrum is in the near infrared area, and has the characteristics of high quantum yield and sensitivity; the erythrocyte and platelet hemolytic agent can rapidly solves erythrocytes in the blood, and is composed of a cationic surfactant, a non-ionic surfactant, a pH buffer agent or combination of the cationic surfactant, the non-ionic surfactant, and the pH buffer agent. The leukocyte classification reagent provided by the invention can carry out precise subgroup identification on lymphocyte, monocyte, neutrophil, basophilic granulocyte, and acidophilic granulocyte in leukocytes of blood samples, and further identify immature cells or abnormal cells.

Description

technical field [0001] The present invention relates to a white blood cell classification and counting kit and a classification method thereof. Specifically, the present invention relates to a kit for classifying and counting white blood cell subgroups on a flow cytometer and using the kit to classify white blood cell subgroups method of counting. Background technique [0002] Leukocytes in human peripheral blood can be divided into five subtypes: monocytes, lymphocytes, eosinophils, neutrophils, and basophils. The proportions of various subtypes of white blood cells in the peripheral blood of normal healthy people are constant. When some diseases occur in the human body, the number of certain types of white blood cells will increase or decrease. For example, neutropenia may occur under symptoms such as acute suppurative infection, granulocytic leukemia, dosage form bleeding, postoperative, heavy metal poisoning, etc. Increased eosinophils may occur in allergies, parasitic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/50G01N33/52G01N21/64
Inventor 邵建辉
Owner 嘉善加斯戴克医疗器械有限公司
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