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Preparation and storage method of chicken erythrocyte

A chicken red blood cell and storage method technology, applied in the field of reagents for medical testing, can solve the problems of inability to remove red blood cells, white blood cells, adhered red blood cells and cell fragments, impurity of chicken red blood cells, non-compliance, etc., so as to reduce the cost of cell preparation and avoid complicated Steps, the effect of reducing the CV value

Inactive Publication Date: 2014-01-29
SUZHOU INST OF BIOMEDICAL ENG & TECH
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Problems solved by technology

However, this method is relatively rough, and cannot remove white blood cells, adhered red blood cells, and cell debris from the red blood cells, resulting in the impurity of the obtained chicken red blood cells. Red blood cell CV value below 10% standard

Method used

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  • Preparation and storage method of chicken erythrocyte
  • Preparation and storage method of chicken erythrocyte
  • Preparation and storage method of chicken erythrocyte

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Embodiment Construction

[0030] The present invention will be described in detail below with reference to the accompanying drawings and in combination with embodiments.

[0031] Unless otherwise specified, the reagents used in the following examples are analytical reagents.

[0032] 1. Chicken red blood cell extraction and purification

[0033] (1) Temporarily store the freshly collected chicken red blood cells anticoagulated with heparin sodium at 4°C, and separate them as soon as possible.

[0034] (2) Mix anticoagulated venous blood with 3% gelatin solution at a ratio of 1:1, and let it stand to separate the solution.

[0035] Preferably, the gelatin solution formula is: 30 g of high-quality gelatin, 0.27 g of potassium dihydrogen phosphate, 1.42 g of disodium hydrogen phosphate, 8 g of sodium chloride and 0.2 g of potassium chloride, dissolved in 1 L of deionized water, heated in a boiling water bath Dissolve and store at 4°C until use.

[0036] (3) Remove the upper layer solution, and resuspen...

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Abstract

The invention discloses a preparation and storage method of chicken erythrocyte. The method comprises the following steps: collecting heparin anticoagulation venous blood of an adult healthy rooster; purifying by adopting the following steps: removing leukocyte by employing a gelatin sedimentation method; removing synechia among cells by ethylenediamine tetraacetic acid treatment; removing soterocyte and cell debris by employing a low-speed centrifugal process; fixing and storing the purified chicken erythrocyte in a glutaraldehyde solution; dyeing regularly and detecting an X-axis half-peak variation coefficient (CV) of chicken erythrocyte deoxyribonucleic acid (DNA) in flow cytometry detection, wherein chicken erythrocyte deoxyribonucleic acid can be used when X-axis half-peak variation coefficient is within 10%. The chicken erythrocyte CV value obtained by the invention accords with the commercially available standard, is simple and convenient to operate, and strong in repeatability, and can be developed as the commercially available product.

Description

[0001] technical field [0002] The invention belongs to the field of reagents for medical detection, and in particular relates to the preparation and storage of chicken red blood cells for calibrating flow cytometers. Background technique [0003] Flow cytometry is a new technology developed in the past two decades. This method is highly sensitive, easy to operate, and can provide various information about cells. Therefore, it is widely used in research in the fields of biology, clinical medicine, and pharmacy. . However, in actual use, flow cytometers need to use calibration reagents for calibration and stability analysis before sample testing can be performed. Chicken erythrocytes are one of the main reagents for the calibration and stability assessment of the coefficient of variation (CV) of flow cytometry. In addition, since the DNA content is 1 / 3 of that of normal human tissue cells, adding chicken red blood cells to human tissue cell samples and co-staining can comp...

Claims

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Application Information

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IPC IPC(8): C12N5/078A01N1/02
Inventor 武晓东殷建王策
Owner SUZHOU INST OF BIOMEDICAL ENG & TECH
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