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A method for separating and identifying nucleated red blood cells in blood

A nuclear red blood cell and blood technology, applied in the field of blood separation, identification and detection, can solve the problems of high cost, cumbersome operation and high technical requirements

Inactive Publication Date: 2011-11-30
孙艳萍
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The direct acquisition method is costly, or the specificity is not strong, easy to be polluted, or the operation is cumbersome, the technical requirements are high, its reliability and practicability are questioned, and it is difficult to popularize and apply

Method used

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  • A method for separating and identifying nucleated red blood cells in blood
  • A method for separating and identifying nucleated red blood cells in blood

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Embodiment Construction

[0090] (1) Preparation of main reagents

[0091] 1. Polylysine working solution: fully mix polylysine stock solution with double distilled water 1:10, and wait for the bubbles to disappear for later use;

[0092] 2. 0.01mol / L PBS (PH 7.4) containing 0.5% BSA

[0093] Sodium chloride 6.5g

[0094] Na 2 HPO 4 12H 2 O 4g

[0095] NaH 2 PO 4 2H 2 O 0.35g

[0096] Add 800ml of double distilled water, adjust the pH value to 7.4, and make up to 1000ml with double distilled water. 0.01mol / L PBS, after autoclaving, add 5g BSA, store at 40°C.

[0097] 3. Alkaline cell lysate (containing KOH 200mmol / L, DTT 50mmol / L)

[0098] 400mmol / L KOH 1ml

[0099] 1mol / L DTT 100ul

[0100] Sterile water for injection is 850-900ul, mixed thoroughly, filtered through a 0.22μm microporous filter, subpackaged, and stored at -20°C for later use.

[0101] (2) Anti-degumming slide preparation

[0102] 1. Pretreatment of slides

[0103] Wash the ordinary glass slide with hot soapy water, clea...

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Abstract

The invention discloses a method for separating and identifying nucleated erythrocytes in blood, which comprises the following steps: (1) preparing reagents; (2) preparing anti-degumming glass slides; Operation method to obtain nucleated red blood cells in the blood. The method for separating and identifying nucleated red blood cells in blood of the present invention can effectively identify nucleated red blood cells, all 40 samples can detect nucleated red blood cells, the detection rate is 100%, and 2×106 to 6×106 nucleated red blood cells were found in each case ml. The present invention is easy to identify nucleated red blood cells after benzidine staining, and can quickly obtain single nucleated red blood cells by skilled micromanipulation, laying a good preliminary foundation for applying single nucleated red blood cells to gene analysis.

Description

technical field [0001] The invention relates to the technical field of blood separation, identification and detection, in particular to a method for separating and identifying nucleated red blood cells in blood. Background technique [0002] Separation of blood samples into various cellular components requires adjustment of blood sample handling / processing techniques accordingly. A conventional method of separating a whole blood sample into its various cellular components is by centrifugation. While effective, this method is labor intensive, relatively inefficient and requires manual control of the cellular fraction of the blood sample. [0003] The separation of nucleated red blood cells is the beginning of genetic analysis, and it is also one of the more difficult steps. Scholars from various countries have tried various methods to separate nucleated red blood cells, but have not found a direct, reliable and efficient way to separate them. [0004] Attempts to carry out...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/49G01N1/30G01N1/34G01N21/25
Inventor 孙艳萍
Owner 孙艳萍
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