Dual-Dye Flow Cytometry for Leukocyte and Infected Erythrocyte Detection
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Solution Overview
Problem
Current methods for detecting malaria parasites require separate tests for leukocyte and infected erythrocyte detection, increasing test time, blood volume, and costs, and existing methods struggle with accurate detection in the presence of reticulocytes.
Innovation Solution
A hematology analyzer uses a combination of flow cytometry and fluorescence staining with two dyes to simultaneously detect leukocytes and infected erythrocytes in a single test, employing a hemolytic agent to lyse erythrocytes while staining leukocytes and infected erythrocytes with specific dyes, and analyzing optical signals to classify and count particles.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If separate tests are used for leukocyte detection and infected erythrocyte detection, then detection accuracy for each parameter is maintained, but test time and blood volume requirements increase
Solution Approach 1:
The patent combines leukocyte detection and infected erythrocyte detection into a single integrated test system. The flow cytometer simultaneously measures multiple parameters (leukocyte count, infected erythrocyte count, reticulocyte count) from one blood sample using multiple detectors and analysis methods, eliminating the need for separate tests while maintaining detection accuracy through advanced signal processing and particle classification algorithms.
Solution Approach 2:
The flow cytometer is designed with multi-functionality to perform both leukocyte detection and infected erythrocyte detection using the same instrument and sample. The system uses multiple light sources, detectors, and analysis modes to achieve universal detection capabilities across different cell types and infection stages, reducing test time and blood volume requirements.
2Measurement precision
If separate tests are used for leukocyte detection and infected erythrocyte detection, then detection accuracy for each parameter is maintained, but blood volume and reagent costs increase
Solution Approach 1:
The patent combines leukocyte detection and infected erythrocyte detection into a single integrated test system. The flow cytometer simultaneously measures multiple parameters (leukocyte count, infected erythrocyte count, reticulocyte count) from one blood sample using multiple detectors and analysis methods, eliminating the need for separate tests while maintaining detection accuracy through advanced signal processing and particle classification algorithms.
3Measurement precision
If fluorescent dyes are used to stain infected erythrocytes under hemolytic conditions, then detection accuracy is improved, but reticulocytes cannot be accurately detected
Solution Approach 1:
The patent segments the detection process into multiple independent analysis channels within the flow cytometer. Different detectors measure different parameters (scattered light, fluorescence) to identify different cell types (leukocytes, infected erythrocytes, reticulocytes) based on their unique optical properties. This segmentation allows simultaneous accurate detection of all cell types even under hemolytic conditions with fluorescent staining.
Solution Approach 2:
The patent adds multiple detection dimensions (forward scattered light, side scattered light, multiple fluorescence channels) to differentiate between cell types. By measuring particles across multiple optical dimensions simultaneously, the system can distinguish infected erythrocytes from reticulocytes and leukocytes even when fluorescent dyes are used, resolving the limitation of previous single-dimension detection methods.
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
Enables simultaneous detection of leukocyte and infected erythrocyte parameters in a single test, reducing blood volume and costs, while maintaining accuracy even in the presence of reticulocytes.
Implementation Method 1
a hemolytic agent to lyse erythrocytes
Implementation Method 2
fluorescence staining with two dyes to simultaneously detect leukocytes and infected erythrocytes
Implementation Method 3
analyzing optical signals to classify and count particles
Data Source
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AI summary
A sample analysis method for analyzing a blood sample, a sample analyzer, and a computer-readable storage medium. Optical signals generated when particles in a test sample solution are illuminated by an excitation light when passing one by one through an optical detection area are acquired in one test, said sample solution being acquired when a blood sample is treated with a hemolytic agent, a first dye, and a second dye, the first dye being capable of dyeing white blood cells, the second dye being capable of dyeing infected red blood cells, the optical signals comprising a scattered light signal, a first fluorescent signal corresponding to the first dye, and a second fluorescent signal corresponding to the second dye; white blood cell optical information is acquired on the basis of the scattered light signal and of the first fluorescent signal; and red blood cell optical information is acquired on the basis of the scattered light signal and of the second fluorescent signal. Implemented by means of the present method is the simultaneous acquisition of the white blood cell optical information and the infected red blood cell optical information in a same detection channel or in a same test, thus reducing the volume of blood used in and costs for testing.