Near-Infrared Fluorescent Dye for Reticulocyte Counting
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Solution Overview
Problem
Current methods for counting reticulocytes in blood analysis, such as visual counting and early flow cytometry, face challenges like long assay times, susceptibility to human error, and interference from background fluorescence, leading to poor repeatability and accuracy.
Innovation Solution
A reagent comprising a fluorescent dye with a specific formula and a surfactant, which rapidly stains cells and reduces orientational noise, allowing for accurate differentiation and counting of reticulocytes using flow cytometry by emitting signals in the near-infrared region, minimizing background interference.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If visual counting or early flow cytometry methods are used for reticulocyte counting, then the assay can be performed, but the assay time is long and repeatability is poor
Solution Approach 1:
The patent changes the fluorescence emission parameter by selecting dyes that emit in the near-infrared region (650-900 nm), which fundamentally alters the detection characteristics and enables both rapid processing and high repeatability through automated flow cytometry
Solution Approach 2:
The patent replaces manual visual counting with automated flow cytometry detection, substituting mechanical/optical manual observation with an automated electronic detection system that provides consistent, repeatable measurements
2Measurement precision
If early fluorescent dyes are used for staining, then reticulocytes can be differentiated, but background fluorescence interferes with accurate counting
Solution Approach 1:
The patent changes the fluorescence emission wavelength parameter from the visible range to the near-infrared region (650-900 nm), which eliminates background fluorescence interference while maintaining accurate reticulocyte differentiation capability
Solution Approach 2:
The patent utilizes fluorescence emission in the near-infrared region instead of visible light, effectively changing the 'color' range to avoid overlap with background fluorescence and enable clear signal differentiation
3Measurement precision
If staining time is extended to improve cell differentiation, then accuracy improves, but the overall assay time increases
Solution Approach 1:
The patent changes the chemical parameters of the staining reagent to enhance staining intensity and speed, allowing accurate cell differentiation to be achieved rapidly without requiring extended incubation periods
Solution Approach 2:
The patent uses composite fluorescent dye formulations that combine multiple components to achieve both rapid staining kinetics and high differentiation accuracy, resolving the trade-off between speed and precision
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
The reagent enables rapid and precise differentiation and counting of reticulocytes, improving the accuracy and reliability of blood analysis by reducing the impact of human error and background fluorescence, thus enhancing the precision of erythrocyte generation evaluation and anemia diagnosis.
Implementation Method 1
a compound having the following general formula I as the fluorescent dye... emitting signals in the near-infrared region
Implementation Method 2
a surfactant... which rapidly stains cells and reduces orientational noise
Data Source
AI summary
The present disclosure provides a reagent for blood analysis which may include: (1) a compound having the general formula I as a fluorescent dye, wherein n, X, R1, R2, R3, R4, R5 and Y− are as defined in the specification; (2) a surfactant selected from cationic surfactants, zwitterionic surfactants and anionic surfactants. The present disclosure also provides a method to perform blood analysis including the following steps of: (a) mixing the blood sample with the reagent for blood analysis disclosed to form a cell suspension; (b) detecting the scattered light signals and fluorescence signals from the cells; and (c) differentiating and counting the cells in the blood in terms of the scattered light signals and fluorescence signals.