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Red blood cell lysis solution and lysis method thereof

A technology for red blood cell lysate and blood samples, used in medicine and biology, red blood cell lysate and its lysis field, can solve the problems of insufficient lysis efficiency, low cell purity, and many types of cells, so as to overcome the inability to stably preserve the remaining cells. Components, safe toxic and side effects, effects without toxic and side effects

Inactive Publication Date: 2014-12-03
SUREXAM BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The tumor cells separated by density gradient centrifugation are impure, the separation efficiency is not high, and a considerable part of the tumor cells are lost. The natural sedimentation method and the differential sedimentation method are simple, but there are many types of cells in each layer, and the purity of the separated cells is not high. The most widely used ammonium chloride separation method also has the problem of poor purity of isolated cells
[0004] At present, many conventional erythrocyte lysing reagents have defects that the lysing effect is not good enough in practical applications, for example, the purity of the isolated cells is poor, or the lysing efficiency is not high enough, etc.

Method used

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  • Red blood cell lysis solution and lysis method thereof
  • Red blood cell lysis solution and lysis method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Embodiment 1. A kind of erythrocyte lysate

[0016] 1, a kind of erythrocyte lysate, has following composition:

[0017] test group

[0018] Distilled water was added to 1L, the concentration of formaldehyde was volume ratio, and the pH was adjusted to 7.4.

[0019] 2. Lysis of red blood cells:

[0020] 1) Take 5ml of blood from the same specimen, and the initial concentration of red blood cells is 5×10 12 / L;

[0021] 2) Add 15ml of erythrocyte lysate reagent respectively, and mix the two thoroughly;

[0022] 3) Place at 4°C for 20 minutes, centrifuge at 600g for 5 minutes, and discard the supernatant. The RBC was counted by an automatic blood cell counter, and the results were as follows:

[0023] Table 1 Average number of red blood cells ( / L)

[0024] sample

[0025] 5

[0026] It can be seen from the above data that after the erythrocyte lysate of the present invention is lysed, the lysates of the five groups can effectively lyse...

Embodiment 2

[0027] Embodiment 2 The effect contrast of red blood cell lysing of different formulas

[0028] 1. Formula of erythrocyte lysate

[0029] According to literature reports, three commonly used erythrocyte lysing formulas were selected for comparison of lysing effects. Use 100ul whole blood, mix according to the volume ratio of whole blood to lysate of 1:3, and set a series of working concentration gradients, as follows:

[0030]

[0031] Recipe 1 (1×): 1M Tris0.1M, NaCl0.1M, MgCl 2 ·6H 2 O0.05M;

[0032] Recipe 2 (1×): NH 4 Cl0.1396M, 1M Tris0.1696M;

[0033] Recipe 3 (1×): NH 4 Cl0.155M, KHCO 3 0.01M, EDTA-Na 2 0.002M, 0.05% formaldehyde.

[0034] Mix according to the ratio of whole blood to lysate 1:3, place at 4°C for 20 minutes, centrifuge the processed sample at 600g for 5 minutes, and count WBC and RBC with an automatic blood cell counter. The conventional evaluation standard in the technical field of the present invention is: the amount of sediment after cent...

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Abstract

The invention provides a red blood cell lysis solution and a method for lysing red blood cells by using the red blood cell lysis solution. The red blood cell lysis solution comprises the following components by concentration: 0.1 M to 0.5 M of ammonium chloride, 0.001 M to 0.2 M of potassium bicarbonate and 0.001 to 0.05 M of ethylenediamine tetraacetic acid or disodium ethylene diamine tetraacetate. The red blood cell lysis solution provided by the invention has the advantages of a good red blood cell lysis effect, safety to and no toxic and side effect on operators and high lysis efficiency. Formaldehyde is used as a fixing agent, and the red blood cell lysis solution is used together with other components to realize removal of red blood cells in blood; moreover, the solution can be used as a protection agent for a part of non-erythrocyte cells in a blood sample for sample transport, thereby overcoming the disadvantage that remaining cell components in purified blood cannot be stably preserved in the prior art.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a erythrocyte lysate and a lysing method thereof. Background technique [0002] Red blood cells, also known as red blood cells (RBC), are the most numerous blood cells in the blood, and are also the most important medium for transporting oxygen through the blood in vertebrates, and also have immune functions. Red blood cells contain hemoglobin, which makes blood red. However, when modern clinical diagnostic methods are used for diagnostic testing of blood samples, red blood cells interfere with many diagnostic tests, which requires the separation of red blood cells from blood samples before further testing of the samples. [0003] At present, the methods for removing red blood cells mainly include density gradient centrifugation, natural sedimentation, differential sedimentation, and ammonium chloride separation. The tumor cells separated ...

Claims

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Application Information

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IPC IPC(8): C12N1/06
Inventor 许嘉森刘志明吴诗扬杨威威
Owner SUREXAM BIO TECH
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