Platelet Analysis Using Oxazine Dye Staining

Resolve Bottlenecks,
Find Innovative Solutions
Generate Solutions

Solution Overview

Problem

Current methods for measuring platelets in blood, such as electrical resistance and antibody-based techniques, face challenges in accuracy and speed, especially when contaminants like lipid particles and fragmented red blood cells are present, making it difficult to rapidly and accurately determine platelet counts in clinical settings.

Innovation Solution

A method using flow cytometry with specific oxazine dyes like Capri blue, Nile blue, and brilliant cresyl blue to stain platelets, allowing for their clear discrimination from other blood cells and contaminants, enabling more accurate and rapid platelet counting by measuring scattered light and fluorescence.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If electrical resistance method is used for platelet measurement, then the measurement can be performed rapidly, but measurement accuracy deteriorates when contaminants are present

Engineering Contradiction:
Improvemeasurement speedVSAvoidplatelet count accuracy
Core Design Contradiction:
ProductivityVSMeasurement precision

Solution Approach 1:

The patent applies fluorescent staining with oxazine dyes that selectively bind to platelets, causing them to emit fluorescence at specific wavelengths. This optical property change enables clear discrimination between platelets and contaminants (lipid particles, fragmented red blood cells) based on their different fluorescence characteristics, thereby maintaining high measurement accuracy while preserving rapid measurement capability

Inventive Principle:
Principle #32Color changes

Solution Approach 2:

The patent changes the detection parameter from electrical resistance to fluorescent light emission. By measuring fluorescence intensity and wavelength characteristics rather than electrical impedance, the system can accurately identify platelets even in the presence of contaminants that would interfere with resistance-based measurements, thus resolving the accuracy-speed contradiction

Inventive Principle:
Principle #35Parameter changes

2Measurement precision

If antibody-based flow cytometry method is used for platelet measurement, then measurement accuracy is improved, but measurement time increases

Engineering Contradiction:
Improveplatelet count accuracyVSAvoidmeasurement time
Core Design Contradiction:
Measurement precisionVSLoss of time

Solution Approach 1:

The patent replaces expensive, time-consuming antibody reagents with simple, rapidly-acting fluorescent dyes (oxazine dyes). These dyes provide sufficient specificity for platelet identification without requiring complex antigen-antibody reactions, thereby maintaining high measurement accuracy while dramatically reducing measurement time to meet urgent clinical needs

Inventive Principle:
Principle #27Cheap short-living objects (Disposable)

Solution Approach 2:

The patent substitutes the biochemical mechanism (antigen-antibody reaction) with a simpler optical mechanism (fluorescent staining). This replacement eliminates the time-consuming immunological reaction step while preserving the ability to specifically identify and count platelets through their unique fluorescence signature

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

3Productivity

If conventional flow cytometry without specific staining is used, then measurement speed is maintained, but ability to discriminate platelets from contaminants deteriorates

Engineering Contradiction:
Improvemeasurement speedVSAvoiddiscrimination accuracy
Core Design Contradiction:
ProductivityVSMeasurement precision

Solution Approach 1:

The patent introduces fluorescent staining that causes platelets to emit light at specific wavelengths distinct from contaminants. Lipid particles and fragmented red blood cells do not exhibit the same fluorescence characteristics as platelets when stained with oxazine dyes, enabling clear visual and computational discrimination while maintaining rapid flow cytometry measurement speed

Inventive Principle:
Principle #32Color changes

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

This approach enhances the accuracy and speed of platelet counting, effectively distinguishing platelets from contaminants and other blood cells, even in samples with low platelet counts, thereby improving clinical diagnostics.

Implementation Method 1

preparing a measurement sample by mixing a sample and a dye for staining platelet, the dye being selected from the group consisting of Capri blue, Nile blue and brilliant cresyl blue

Methodology Applied
Scientific EffectAbsorption (physical): Absorption (physical)

Implementation Method 2

measuring scattered light and fluorescence emitted from cells in the measurement sample by irradiating the cells with light

Methodology Applied
Scientific EffectScattering: Scattering

Implementation Method 3

measuring scattered light and fluorescence emitted from cells in the measurement sample by irradiating the cells with light

Methodology Applied
Scientific EffectFluorescence: Fluorescence

Data Source

PatentEP1918709B1Analyzing method for platelets
Publication Date: 2015.04.01 SYSMEX CORP
  • EP1918709B1 patent drawingFigure 1A
  • EP1918709B1 patent drawingFigure 1B
  • EP1918709B1 patent drawingFigure 1C

AI summary

A method for analyzing platelet is described. In the method, a measurement sample is prepared by mixing a sample and a dye for staining platelet. The dye is selected from the group consisting of Capri blue, Nile blue and brilliant cresyl blue. By irradiating cells in the measurement sample with light, scattered light and fluorescence emitted from the cells is measured. The platelet is detected on the basis of the scattered light and the fluorescence. A reagent kite and a reagent are also described.