Hemolytic Reagent Composition for Accurate White Blood Cell Classification
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Solution Overview
Problem
Existing hemolytic reagents struggle to accurately classify white blood cells into lymphocyte, monocyte, neutrophil, and eosinophil populations.
Innovation Solution
A hemolytic reagent containing a nonionic surfactant of specific formula (R1—R2—(CH2CH2O)n—H) with controlled concentration and a staining reagent for nucleic acid fluorescence, used in an automatic hemocytometer for flow cytometry, to enhance classification accuracy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional hemolytic reagents are used, then red blood cells are hemolyzed, but white blood cell classification accuracy is insufficient
Solution Approach 1:
The patent changes the chemical parameters of the hemolytic reagent by specifying a nonionic surfactant with a particular molecular structure (formula I) where R1 is an alkyl/alkenyl/alkynyl group with 8-25 carbon atoms, R2 is oxygen or carboxyl group, and n is 23-30. The concentration is precisely controlled at 1700-2300 ppm, which optimizes the hemolysis effect while preserving white blood cell integrity for accurate classification into five types.
Solution Approach 2:
The patent creates a composite reagent system combining the nonionic surfactant (formula I) with a staining reagent that fluorescently stains nucleic acid. This composite approach allows simultaneous hemolysis of red blood cells and fluorescent staining of white blood cell nuclei, enabling accurate flow cytometry-based classification of all five white blood cell types.
2Quantity of substance
If hemolytic reagent concentration is increased to improve hemolysis, then red blood cell lysis improves, but white blood cell damage increases
Solution Approach 1:
The patent optimizes the concentration parameter of the nonionic surfactant to a specific range of 1700-2300 ppm, which is sufficient to achieve complete red blood cell hemolysis while maintaining white blood cell membrane integrity. This precise parameter control resolves the contradiction between hemolysis efficiency and white blood cell protection.
Solution Approach 2:
The nonionic surfactant with the specific molecular structure (formula I) exhibits selective action: it effectively disrupts red blood cell membranes while having minimal harmful effect on white blood cell membranes. This local quality difference in membrane interaction allows differential hemolysis at the optimized concentration range.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The reagent kit achieves precise classification of lymphocyte, monocyte, neutrophil, and eosinophil populations, improving differentiation and reducing dispersion in optical information.
Implementation Method 1
a nonionic surfactant represented by formula (I) below: R1—R2—(CH2CH2O)n—H
Implementation Method 2
a staining reagent that fluorescently stains nucleic acid
Data Source
AI summary
Disclosed is a hemolytic reagent comprising a nonionic surfactant represented by formula (I) below:R1—R2—(CH2CH2O)n—H (I)where R1 represents an alkyl group, an alkenyl group, or an alkynyl group having 8 or more and 25 or less carbon atoms,R2 represents an oxygen atom, (COO) or a group represented by formula (II) below:where n is 23 or larger and 25 or smaller, or 30,with n of 23 or larger and 25 or smaller, a concentration of the nonionic surfactant is 1700 ppm or higher and 2300 ppm or lower, andwith n of 30, the concentration of the nonionic surfactant is 1900 ppm or higher and 2300 ppm or lower.