Method and apparatus for measuring hematological sample

A technology for measuring devices and blood samples, applied in measuring devices, immunoassays, biological tests, etc., can solve the problems that appear in the area of ​​white blood cells and red blood cells, and may appear in the area of ​​bone marrow blasts in (3), and achieve Exclude platelet aggregation, accurate measurement effect

Active Publication Date: 2007-10-03
SYSMEX CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, according to the technique described in WO2004 / 001408, although bone marrow blasts can be classified in (3), if a sample containing a small amount of platelet aggregation is measured, platelet aggregation may occur in (1) step Areas of white and red blood cell ghosts, which may appear in (3) in areas of bone marrow blasts

Method used

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  • Method and apparatus for measuring hematological sample
  • Method and apparatus for measuring hematological sample
  • Method and apparatus for measuring hematological sample

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0100] [Preparation of samples for measurement]

[0101] A sample for measurement is prepared by mixing a solution containing the treatment reagent and a blood sample. If the above-mentioned kit is used, a solution containing a surfactant (hemolyzing agent) is first mixed with a blood sample, and then a dye is mixed to prepare a sample for measurement.

[0102] When blood sample and treatment reagent are mixed, the ratio of sample: treatment reagent is 1:10-1:1000, the reaction temperature is 20-40°C, and the reaction time is 3 seconds to 5 minutes, preferably 4 seconds to 1 minute.

[0103] In the measurement sample thus prepared, erythrocytes and mature leukocytes shrink smaller. In particular, red blood cells have no nuclei, and the hemolysis and cell shrinkage caused by damaged cell membranes are more serious, and the fluorescent pigment cannot be stained. Therefore, the forward scattered light, side scattered light and fluorescence are all weakened, which can be treated ...

experiment example

[0185] [blood sample]

[0186] The blood samples used were a sample containing a large amount of bone marrow blasts (sample A), a sample containing bone marrow blasts and immature leukocytes (sample B). Samples A and B contain platelet aggregation.

[0187] [Preparation of samples for measurement]

[0188] (1) Preparation of a measurement sample using the treatment reagent A

[0189] (A-1) Hemolytic agent

[0190] A hemolytic agent of the following structure containing the following components was prepared.

[0191] Glycine 19.5g / l

[0192] Polyoxyethylene(16) oleyl ether 25.0g / l

[0193] Sodium N-lauroyl sarcosinate 0.75g / l

[0194] HEPES 10mM

[0195] distilled water 11

[0196] Caustic soda pH up to 7.0

[0197] (A-2) Dyeing agent

[0198] Fluorescent dye A represented by the following formula was dissolved in ethylene glycol.

[0199]

[0200] The hemolytic agent and the blood sample are mixed, and the dyeing agent is added thereto until the final concentrati...

experiment example 1

[0211] Mix blood sample A and processing reagent B, react at 33°C for 5 seconds, prepare a sample, put the sample into a flow cell, and use a flow cytometer (light source: red semiconductor: wavelength: 633nm) to measure forward scattered light, side Scattered light and fluorescence. The measurement results were plotted as the first scatter plot with the forward scattered light intensity and the side scattered light intensity as the two axes (Fig. 13(A)) and the second scatter plot with the fluorescence intensity and the forward scattered light intensity as the two axes Figure (Fig. 13(B)).

[0212] In FIG. 13(A), the region where the first cell population appears is the solid line region. In FIG. 13(B), the region where the second cell population appears is the solid line region. In FIG. 13(A) , the expanded cell population containing lymphocytes partially overlaps with the first cell population.

[0213] The cell population contained in both the first cell population and ...

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Abstract

The invention provides a measuring method which can classify and count myeloblast more precisely without influence of other component in a sample including platelet aggregation in measurement of a blood sample with a flowcytometry, wherein damage is given to a cell membrane of erythrocyte and mature leukocyte contained in a hematological sample, a hemocyte in which a cell membrane is damaged is constricted, and this is dyeing-treated with a fluorescent dye which can stain a nucleic acid to obtain a sample, the sample is measured with a flowcytometer, and a cell contained in a first cell group containing myeloblast, which is specified based on forward scattered light information and side scattered light information, and contained in a second cell group containing myeloblast, which is specified based on forward scattered light information and fluorescent information, is counted as myeloblast. The invention further provides a measuring device comprising following systems: a sample treating system, an information acquisition system, a first specific system, a second specific system and a counting system.

Description

Technical field: [0001] The invention relates to a blood sample measuring method and a measuring device, which can accurately distinguish and count immature white blood cells, especially bone marrow blast cells, from other blood cells. Background technique: [0002] The cellular components of blood are roughly divided into platelets, white blood cells and red blood cells, and white blood cells are further divided into granulocytes (eosinophils, neutrophils and basophils), monocytes and lymphocytes. These cells are produced in the bone marrow, differentiate from immature cells, mature, and move to the peripheral blood. Immature white blood cells (immature white blood cells) that have not developed into white blood cells in normal blood such as granulocytes, monocytes and lymphocytes (hereinafter sometimes referred to as "mature white blood cells") will not appear in the peripheral blood of normal healthy people, but leukemia, Immature white blood cells such as bone marrow bl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N15/12G01N33/48
CPCG01N15/1456Y10T436/25125G01N2015/1402G01N15/1459Y10T436/25G01N2015/1006Y10T436/101666G01N2015/1477Y10T436/10
Inventor 辻智悠吉田步小国振一郎
Owner SYSMEX CORP
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