Flow Cytometry Myeloblast Classification via Hemolysis and Fluorescence
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Solution Overview
Problem
Current methods for measuring hematological samples using flowcytometry face challenges in precisely classifying and counting myeloblasts, especially when platelet aggregation is present, leading to potential misclassification and reduced precision.
Innovation Solution
A method and apparatus that utilize a treating reagent to damage erythrocyte and mature leukocyte cell membranes, allowing for differential staining and separation of myeloblasts from other cells by exploiting differences in membrane damage and fluorescent dye penetration, using a combination of surfactants and fluorescent dyes to enhance the distinction in flow cytometry analysis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Extent of automation
If flowcytometry is used to measure hematological samples, then automated measurement of blood cells is achieved, but measurement precision deteriorates when platelet aggregation is present
Solution Approach 1:
The patent applies preliminary action by treating the blood sample with a hemolytic agent before flowcytometry measurement to lyse red blood cells and prevent platelet aggregation. This preliminary treatment eliminates the interfering factors (RBCs and platelet aggregates) that would otherwise compromise myeloblast detection accuracy, thereby enabling precise automated measurement without sacrificing automation.
Solution Approach 2:
The patent converts the harmful effect of platelet aggregation into a beneficial discrimination criterion. By using scattergram analysis, the method leverages the different light scattering properties of platelet aggregates versus myeloblasts to selectively identify and count myeloblasts. The previously harmful aggregation artifact becomes a useful distinguishing feature that enhances measurement precision.
2Measurement precision
If multiple-dimensional scattergram analysis is used to classify cell types, then cell classification capability is improved, but device complexity increases
Solution Approach 1:
The patent applies segmentation by dividing the cell classification process into multiple sequential two-dimensional scattergram analyses rather than using a single complex multi-dimensional system. Each scattergram focuses on specific parameters (e.g., forward scatter vs. side scatter, fluorescent intensity vs. scatter) to progressively narrow down cell types. This segmented approach achieves high classification precision while keeping each individual analysis step relatively simple.
Solution Approach 2:
The patent utilizes dimensionality change by transitioning from traditional two-dimensional scattergram analysis to three-dimensional analysis incorporating fluorescent intensity as a third dimension. This additional dimension provides further discrimination capability for identifying myeloblasts versus other cell types, enhancing classification precision without requiring overly complex processing algorithms.
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 precise classification and counting of myeloblasts by effectively excluding the influence of platelet aggregation and distinguishing myeloblasts from mature leukocytes, thereby improving measurement accuracy in hematological samples.
Implementation Method 1
The treating reagent contains a surfactant which gives damage to a cell membrane of an erythrocyte and a mature leukocyte
Implementation Method 2
a fluorescent dye which can stain a nucleic acid; obtaining fluorescent intensity
Implementation Method 3
irradiating a cell passing through a flow cell with excitation light of a dye, obtaining information regarding scattered light or fluorescent light emitted from individual cells
Data Source
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AI summary
A measuring method and a measuring apparatus 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.