Neutrophil Extracellular Trap Detection via Fluorescence Intensity
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for detecting neutrophil extracellular traps have low sensitivity, particularly in distinguishing between neutrophils that have and have not released these traps, leading to difficulties in forming independent clusters on scattergrams.
Innovation Solution
A specimen analyzing method using a nucleic acid staining fluorescent dye to stain nucleic acids in neutrophils, combined with osmotic pressure regulation and surfactants, to enhance the visibility of neutrophil extracellular traps by differentiating their fluorescence intensity from that of neutrophils that have released them.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional flow cytometry methods are used to detect neutrophil extracellular traps, then the detection process can be performed, but the sensitivity for separating neutrophil extracellular traps from neutrophils is low and independent clusters cannot be formed on scattergrams
Solution Approach 1:
The patent divides the detection process into two distinct stages: first detecting neutrophils based on scattered light intensity, then detecting neutrophil extracellular traps based on fluorescence intensity. This segmentation allows each detection step to focus on specific characteristics, forming independent clusters on scattergrams and significantly improving detection sensitivity without requiring complex integrated systems
Solution Approach 2:
The patent introduces a nucleic acid staining fluorescent dye as an intermediary substance that specifically stains neutrophil extracellular traps. This intermediary enables optical detection of the traps through fluorescence, creating a clear distinction between traps and neutrophils on scattergrams while maintaining relatively simple detection equipment
2Measurement precision
If conventional staining methods are used, then neutrophils can be detected, but neutrophil extracellular traps cannot be clearly distinguished from neutrophils that have not released traps
Solution Approach 1:
The patent employs fluorescence staining that causes neutrophil extracellular traps to emit light at specific wavelengths when irradiated. This color/亮度 change creates a clear visual and measurable distinction between traps (fluorescent) and intact neutrophils (non-fluorescent or differently fluorescent), enabling accurate differentiation and cluster formation on scattergrams
Solution Approach 2:
The patent adds fluorescence intensity as a new detection dimension beyond scattered light intensity. By plotting both parameters on scattergrams, neutrophils and neutrophil extracellular traps form separate clusters in the two-dimensional space, making differentiation straightforward and improving detection accuracy without increasing measurement complexity
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 method allows for accurate detection and counting of neutrophil extracellular traps with improved sensitivity, enabling timely clinical interventions such as suppressing excessive inflammation.
Implementation Method 1
a step (A) of preparing a measurement specimen from a biological specimen containing blood cells by staining, with a nucleic acid staining fluorescent dye, nucleic acids contained in neutrophils not having released neutrophil extracellular traps
Implementation Method 2
combined with osmotic pressure regulation and surfactants, to enhance the visibility of neutrophil extracellular traps
Implementation Method 3
combined with osmotic pressure regulation and surfactants, to enhance the visibility of neutrophil extracellular traps
Data Source
Figure 1
Figure 2
Figure 3
AI summary
A specimen analyzing method of an embodiment includes: preparing a measurement specimen from a biological specimen containing blood cells by staining, with a nucleic acid staining fluorescent dye, nucleic acids contained in neutrophils not having released neutrophil extracellular traps; obtaining optical information including fluorescence information by irradiating the measurement specimen with light; and detecting, as neutrophil extracellular traps on the basis of the optical information, particles having lower fluorescence intensity than fluorescence intensity obtained from the neutrophils not having released neutrophil extracellular traps.