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

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

[0031] Unless otherwise specified, the reagents used in the following examples are all analytically pure reagents.

[0032] 1. Extraction and purification of chicken red blood cells

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

[0034] (2) Mix the anticoagulated venous blood with 3% gelatin solution in a ratio of 1:1, and let the solution stand for layers.

[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,...

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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...

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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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