前苏联专利SU753346A3 Method of preparing cell suspension

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专利摘要:
Procedures are disclosed for obtaining white blood cell counts and platelet counts by light scattering measurements in which anticoagulated blood samples are treated such that the morphology of the white cells (leukocytes) and platelets remain intact and permit the discrimination of platelets from white cells and of most classes of white cells from one another and their counting without the need for staining.
公开号:SU753346A3
申请号:SU782639945
申请日:1978-07-04
公开日:1980-07-30
发明作者:Ким Ран
申请人:Текникон Инструментс Корпорейшн (Фирма)；
IPC主号:

专利说明:

The suspension is carried out using Od5% acetic acid salt solution. This improves the ratio of platelet signals | B noise, a clear separation of platelets and similar red cells and leukocytes, and good, well-preserved original individual platelet volumes. It is possible, and in some cases, it is preferable to partially dilute the sample by adding an isotonic saline solution, p in front of the detergent treatment stage.
To determine white blood cells, the sample is diluted by adding an isotopic saline solution, preferably after the imcubadia stage. The sample can also be diluted before the processing stage with a detergent. Diluted suspension of white blood cells preserves the morphology of white cells, allowing the unpainted white cells to be separated using light scattering under small or large angles, and makes it possible to differentiate lymphocytes, monocytes, neutrophils and eosinophils without staining.
Example 1. To an anticoagulated whole blood sample (0.1 ml) was added 0.32 L {l of a 1% polyoxyethylene 20 sorbitan monolaurate solution (in isothermic Saline), and the mixture was allowed to incubate at room temperature in for 1 min
A phosphate-buffered formaldehyde solution (0.42 ml) containing 8 vol.% Was added to the mixture with stirring. % form aldehyde a, and incubate the mixture for 2 min at 57 ° C. Determination of white blood cells is carried out by adding 1.2 ml of isotonic saline solution to the incubated mixture, and using a light scattering solution. white blood cells.
To determine the platelets, 1.2 ml of isotonic saline was added to the incubate; above the mixture. Add 0.1 ml of the mixture to 4.0 ml of 0.5% acetic acid-isotonic saline solution. Plate counting and measuring their size were carried out using the light scattering.
Example 2. Diluted with 0.0. ml of anti-coagulated capillary whole blood in the ratio l: lb with isotonic saline, and 0.1 g of the mixture obtained was added to 0.32 1 1g 0.125% polyoxyethylene 20 sorbant monolaurate (in isotonic saline). The mixture is incubated for 1 minute at room temperature, and 0.42 ml of phosphate-buffered formaldehyde solution containing about 8 vol. Is added to it with stirring. % formaldehyde, and incubated at 57 ° C for 2 min.
Differentiation; the white blood cell count in the above suspension is performed using the cBCTOip section.
The determination of trO | M1bots.it is carried out by adding 1.2 ml of isotonic saline solution to the incubated above mixture, 0.1 ml of the mixture obtained is added to 1.0 ml of 0.5% acetic acid solution — isotonic salt solution . Plate counting and measurement of their size are carried out using the light scattering.
The proposed method allows differentiation of platelets from leukocytes and erythrocytes.
F o .rm lu inventions
A method of obtaining a suspension of cells by treating Yurovi with formaldehyde, adding a hemolyzing agent, characterized in that, in order to enable platelet differentiation from leukocytes and erythrocytes, the blood is treated with a detergent, such as polyoxyethylene 20 sorbitol and mono-uraurate, in the blood sample and blood sample is treated with a detergent, such as polyoxyethylene 20 sorbitol and mono-urauyrate, the blood is treated with a detergent, such as polyoxyethylene and 20 sorbitol and mono-urauyrate in the blood. , 0% relative to the mixture, at a pH of from 6 to 8, followed by incubation of the mixture at a temperature of from 56 ° C to 62 ° C for 2 liters.
A source of information,
taken into account when examining:
1. US patent No. 3741875,
cl. C 12 P 1/04, publ. 1973.

权利要求:
Claims (1)
[1]
Claim
A method of obtaining a suspension of cells by treating the blood with formaldehyde, adding a hemolytic agent, characterized in that, in order to allow the differentiation of platelets from leukocytes and red blood cells, the blood is treated with a detergent, such as polyoxyethylene 20 sorbitol monolaurate, from 0.3 to 2.0% with respect to the mixture, at a pH of 6 to 8, followed by incubation of the 45 mixture at a temperature of 55 ° C to 62 ° C for 2 minutes.
Source of information taken into account during the examination:
50 1. US patent No. 3741875, CL. C 12 P
1/04, publ. 1973.

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同族专利:

公开号 | 公开日

SE446911B|1986-10-13|

AU3740078A|1980-01-03|

IT1111407B|1986-01-13|

FR2398305B1|1983-11-18|

BE868071A|1978-12-13|

US4099917A|1978-07-11|

CA1085280A|1980-09-09|

NL7803424A|1979-01-23|

SE7801766L|1979-01-22|

FR2398305A1|1979-02-16|

IT7868082D0|1978-05-11|

DE2830524C2|1987-11-12|

DE2830524A1|1979-02-01|

GB1597638A|1981-09-09|

AU515945B2|1981-05-07|

CH640948A5|1984-01-31|

JPS5422891A|1979-02-21|

引用文献:

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US4040785A|1976-10-18|1977-08-09|Technicon Instruments Corporation|Lysable blood preservative composition|AT390T|1978-11-01|1981-11-15|Contraves Ag|METHOD FOR REDUCING THE INTERFERENCE OF A PHOTOMETRIC MEASUREMENT BY LIGHT DISTRIBUTION IN A SUSPENSION TO BE MEASURED AND REAGENT FOR CARRYING OUT THE METHOD.|

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US4528274A|1982-07-06|1985-07-09|Coulter Electronics, Inc.|Multi-purpose blood diluent and lysing agent for differential determination of lymphoid-myeloid population of leukocytes|

US5045472A|1981-06-26|1991-09-03|Technicon Instruments Corporation|Reagent mixture and composition for treating red blood cells to effect sphering|

US4465774A|1981-07-16|1984-08-14|Sherwood Medical Company|Standard or control material for glysocylated or total hemoglobin determination|

US4637986A|1983-08-22|1987-01-20|Ortho Diagnostic Systems, Inc.|Flow cytometry lysing reagent with leukoprotective agent for producing a 3-part WBC count|

US4704364A|1984-05-18|1987-11-03|Coulter Electronics, Inc.|Hematology control compositions for three populations of leukocytes; and methods for their preparation and use in whole blood control systems|

JPS61502280A|1984-05-31|1986-10-09|

US4751179A|1984-05-31|1988-06-14|Coulter Electronics, Inc.|Method and reagents for differential determination of four populations of leukocytes in blood|

US4745071A|1985-09-05|1988-05-17|Sequoia-Turner Corporation|Method for the volumetric differentiation of blood cells types|

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法律状态:

优先权:

申请号 | 申请日 | 专利标题

US05/817,595|US4099917A|1977-07-21|1977-07-21|Process for preparing a cell suspension from blood for discrimination of white blood cells and platelets from other blood particles|

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