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Nucleated red blood cell simulation particle and blood quality control substance and preparation methods and application thereof

A technology for simulating particles and nucleated red blood cells, which is applied in the quality control of blood cell analyzers and the field of nucleated red blood cell simulation particles. It can solve the problems of large differences in the shape of nucleated red blood cells, high cost of biological macromolecules, and complex processes, etc., to facilitate industrial production. , low cost, simple steps

Active Publication Date: 2011-06-29
SHENZHEN MINDRAY BIO MEDICAL ELECTRONICS CO LTD
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  • Application Information

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Problems solved by technology

The disadvantage of this method is that the process is complex and the cost of biomacromolecules as raw materials is relatively high
The disadvantage of this method is that the erythrocytes of most birds, reptiles, and fish are oval in shape, which is quite different from the nearly round shape of human nucleated erythrocytes
Therefore, in some cases, the nucleated red blood cells of these animals do not well mimic the characteristics of human nucleated red blood cells

Method used

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  • Nucleated red blood cell simulation particle and blood quality control substance and preparation methods and application thereof
  • Nucleated red blood cell simulation particle and blood quality control substance and preparation methods and application thereof
  • Nucleated red blood cell simulation particle and blood quality control substance and preparation methods and application thereof

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preparation example Construction

[0037] Preparation method of nucleated red blood cell simulation particles

[0038] One embodiment of the present invention provides a method for preparing nucleated red blood cell simulation particles, comprising the following steps:

[0039] Wash and separate anticoagulated blood to obtain purified leukocytes, or further purify leukocytes to obtain leukocyte subsets, such as granulocytes or lymphocytes;

[0040] Suspending leukocytes or leukocyte subsets in a cell treatment solution containing a fluorescent staining inhibitor, the nucleus or nucleic acid of the cell is stably bound to the inhibitor;

[0041] The resulting product is washed.

[0042] The inhibitor is selected from non-fluorescent dyes, fluorescent dyes with lower fluorescence quantum yields than the fluorescent dyes for detection, fluorescent dyes with a large difference in fluorescence spectrum from the fluorescent dyes for detection, and compounds capable of combining with nucleosides.

[0043] There ar...

Embodiment 1

[0077] Take fresh bovine anticoagulant blood and dissolve red blood cells with 0.9% ammonium chloride solution. Bovine white blood cells were separated by centrifugation at 3000 rpm, and the supernatant (containing red blood cell fragments, hemoglobin, etc.) was discarded. Suspend the pelleted leukocytes with PBS and adjust the cell density to about 1×10 10 pcs / liter. Take 10 ml of the above white blood cell suspension, add 1 ml of toluidine blue O staining solution, and mix well. Stain at 30°C for 30 minutes. Stained leukocytes were separated by centrifugation at 3000 rpm, washed with PBS and resuspended.

[0078] Adjust the cell density to about 10 10 cells / L, respectively, adding appropriate concentration of formaldehyde to fix the cells. Fixed leukocytes were separated by centrifugation at 3000 rpm, washed with PBS and resuspended. The fixed cells were detected by using the reagent and method of Example 1 of the Chinese invention patent application with application n...

Embodiment 2

[0083] Take fresh porcine anticoagulant blood and let it stand for a period of time to precipitate red blood cells. Aspirate the supernatant rich in white blood cells, centrifuge at 3000 rpm to separate the cells, and discard the supernatant. Lyse the red blood cells by resuspending the pelleted red blood cells and white blood cells with 0.9% ammonium chloride solution. Separation of porcine leukocytes by centrifugation at 3000 rpm, discarding the supernatant (containing erythrocyte fragments, hemoglobin, etc.) and suspending the precipitated leukocytes with PBS. Use lymphocyte separation medium to separate porcine granulocytes according to the product instructions. Pig granulocytes were washed with PBS. Then the porcine granulocyte suspension was centrifuged (1200 rpm), the supernatant was discarded, and the porcine granulocyte pellet was retained. Add 10ml of Wright's staining solution to the pig granulocyte pellet, pipette the dyeing solution to suspend the pig granulocy...

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Abstract

The invention provides a nucleated red blood cell simulation particle and a preparation method thereof. The particle is a white blood cell combined with a fluorescent dyeing inhibitor, and the inhibitor is stably combined with a nucleus or nucleic acid in the cell, so that the capacity of the particle of combining with a fluorescent dye during detection is reduced. The method comprises the following steps of: (a) obtaining the washed and purified white blood cell; (b) suspending the white blood cell in fluorescent dyeing inhibitor-containing cell processing liquid, wherein the inhibitor is stably combined with the nucleus or the nucleic acid in the cell; and (c) washing the obtained product. The invention also provides a nucleated red blood cell simulation particle-containing blood quality control substance, and application of the nucleated red blood cell simulation particle and the blood quality control substance thereof to the quality control of a blood cell analyzer.

Description

technical field [0001] The invention relates to a blood analysis quality control substance. Specifically, the present invention relates to nucleated red blood cell simulated particles, blood quality control substances containing the nucleated red blood cell simulated particles, and their preparation methods, and the nucleated red blood cell simulated particles and quality control substances can be used in fluorescence-scattering The optical method is used in the quality control of blood cell analyzers based on the detection principle. Background technique [0002] Normal mature human red blood cells do not contain a nucleus. However, in special cases, including certain pathological conditions, nucleated red blood cells (NRBC, also known as erythroblast), a kind of immature red blood cells, will appear in human peripheral blood. The detection of nucleated red blood cells can provide a basis for the diagnosis of certain diseases. Some high-end blood cell analyzers are equip...

Claims

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Application Information

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IPC IPC(8): G01N1/30G01N21/64G01N33/96
CPCG01N2800/22G01N33/96G01N21/6428G01N33/80Y10T436/101666G01N33/5094
Inventor 邓橙徐祖越刘鹤
Owner SHENZHEN MINDRAY BIO MEDICAL ELECTRONICS CO LTD
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