Flow Cytometer Light Scattering for Dye-Free Leukocyte Counting
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Solution Overview
Problem
The counting of nucleated cells is affected by lipid particles when a fluorescent dye is not used in the measurement sample.
Innovation Solution
A measurement method and device that utilize light scattering features to distinguish and count lipid particles, allowing accurate counting of leukocytes without a fluorescent dye by using a flow cytometer to obtain information on lipid particles based on multiple light scattering parameters.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If a fluorescent dye is not used in the measurement sample, then the measurement procedure is simplified and cost is reduced, but lipid particles cannot be distinguished from nucleated cells leading to inaccurate counting
Solution Approach 1:
The patent changes the measurement parameters from fluorescent intensity-based differentiation to light scattering intensity-based differentiation. By measuring forward scattered light intensity (FSC) and side scattered light intensity (SSC) instead of fluorescent dye binding, the system achieves particle differentiation without requiring fluorescent dyes, thus resolving the contradiction between procedure simplicity and counting accuracy
Solution Approach 2:
The patent replaces the chemical staining method (fluorescent dye binding) with an optical scattering method (light scattering intensity measurement). This substitution allows the system to distinguish lipid particles from nucleated cells based on their different light scattering properties rather than chemical affinity, enabling accurate counting without fluorescent dyes
2Measurement precision
If light scattering parameters are used to distinguish lipid particles, then accurate counting is achieved without fluorescent dyes, but the device complexity increases due to multiple optical detection channels
Solution Approach 1:
The patent makes the flow cytometer multi-functional by enabling it to perform both nucleated cell counting and lipid particle detection using the same optical detection system. The single optical detection unit measures multiple light scattering parameters (FSC, SSC) that serve dual purposes: identifying nucleated cells and detecting lipid particles, thus achieving accurate differentiation without adding separate detection systems
Solution Approach 2:
The patent segments the particle identification process into distinct stages: first identifying particles based on forward scattered light intensity (size), then further differentiating based on side scattered light intensity (internal structure). This segmentation of the measurement process allows complex particle differentiation to be achieved through sequential, manageable measurement steps rather than requiring a single complex detection mechanism
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 accurate measurement of leukocyte counts in samples containing lipid particles by distinguishing and correcting for lipid interference, providing reliable results without fluorescent dyes.
Implementation Method 1
obtaining information related to lipid particles contained in the measurement sample based on a plurality of feature values related to light scattering from each particle
Data Source
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AI summary
A measurement method, a measurement device, and a measurement program for acquiring information related to lipid particles contained in a measurement sample prepared without using a fluorescent dye are provided The problem is resolved by the measurement method for measuring the number of particles in a measurement sample prepared without using a fluorescent dye, the method including obtaining information related to lipid particles contained in the measurement sample based on a plurality of characteristic values regarding light scattering in each particle obtained by a flow cytometer from the individual particles contained in the measurement sample.