Fluorescent Dye Composition for Leukocyte Classification in Microfluidics
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for leukocyte differential counting are not optimized for miniaturization, requiring complex systems with both light scattering and fluorescence detection, making them unsuitable for miniaturized instruments like microfluidic devices used in spaceflight and bedside healthcare applications.
Innovation Solution
A diagnostic composition comprising a first cationic dye, a second dye with less cationic charge, and a third anionic dye reactive to proteins, used for fluorescence-based leukocyte differentiation in microfluidic cytometers, allowing for the classification of various leukocyte types through fluorescence detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If light scattering detection and fluorescence detection are combined for leukocyte classification, then classification accuracy is improved, but system complexity increases
Solution Approach 1:
The patent extracts and eliminates the light scattering detection component from the traditional two-method system, retaining only fluorescence detection. This is achieved by using fluorescent dyes that selectively bind to different leukocyte types, allowing classification based solely on fluorescence emission patterns, thereby simplifying the system while maintaining diagnostic capability
Solution Approach 2:
The patent changes the detection parameter from a combination of light scattering and fluorescence to fluorescence-only detection. By using multiple fluorescent dyes with different excitation and emission characteristics that selectively bind to specific leukocyte types, the system achieves classification accuracy through fluorescence parameter variations alone, eliminating the need for light scattering detection
2Measurement precision
If multiple detection steps are used for leukocyte analysis, then classification accuracy is improved, but ease of operation deteriorates
Solution Approach 1:
The patent merges multiple detection steps into a single fluorescence detection step by using a combination of fluorescent dyes that simultaneously stain different leukocyte types. This allows all necessary classification information to be obtained in one measurement process, making the method suitable for miniaturized instruments with limited operational complexity
Solution Approach 2:
The fluorescent dye mixture serves multiple functions simultaneously: it stains different leukocyte types with distinct fluorescence patterns, enables single-step detection, and provides classification information for all major leukocyte subtypes through a unified detection approach, making it universally applicable to miniaturized flow cytometers
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 efficient classification and enumeration of leukocyte subpopulations, such as lymphocytes, monocytes, neutrophils, eosinophils, and basophils, using fluorescence detection in microfluidic devices, improving point-of-care diagnostics and reducing system complexity.
Implementation Method 1
The present invention provides a diagnostic composition, comprising, consisting essentially of, or consisting of: a first dye having a cationic charge; a second dye having less of a cationic charge compared to the first dye; and a third dye being reactive to a protein, having an affinity to a protein or having an anionic charge
Data Source
AI summary
The present invention provide compositions and methods for classifying leukocytes in a leukocyte population using fluorescence detection. The methods include contacting a leukocyte population in a sample having one or more leukocyte types with a diagnostic composition, exciting the leukocyte population with a light source; and measuring emitted light from each of the one or more leukocyte types to classify the leukocyte population.