Aminocoumarin Fluorescent Dye for Reduced Background Interference
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Solution Overview
Problem
Conventional sulfonated coumarin-based dyes have excitation spectra maximized at wavelengths of 470 nm or less, leading to reduced excitation light absorption at 475 nm or more, resulting in high background fluorescence and low signal-to-background ratios during immunofluorescence detection, especially when multiple staining is performed.
Innovation Solution
An aminocoumarin compound with a specific structure, represented by formula (1), is developed, which has an excitation spectrum maximum wavelength of 475 to 510 nm and a fluorescence spectrum maximum wavelength of 510 to 540 nm, allowing for improved fluorescence detection with reduced background noise.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If conventional sulfonated coumarin-based dyes are used, then water solubility is improved, but excitation light absorption is reduced at wavelengths of 475 nm or more
Solution Approach 1:
The patent modifies the chemical structure of coumarin dyes by introducing amino groups at specific positions (formula (1) with R1-R12 representing hydrogen, methyl, or hydroxyl groups). This structural parameter change shifts the excitation spectrum maximum to longer wavelengths (475-510 nm) while maintaining water solubility through sulfonate groups, thereby resolving the contradiction between solubility and excitation absorption at longer wavelengths
Solution Approach 2:
The invention creates composite fluorescent dyes combining coumarin core structure with amino substituents and sulfonate groups. This composite molecular design integrates multiple functional characteristics: the coumarin backbone provides fluorescence, amino groups extend excitation wavelength, and sulfonate groups ensure water solubility, simultaneously achieving all desired properties
2Illumination intensity
If conventional sulfonated coumarin-based dyes are used, then green fluorescence emission is achieved, but background fluorescence increases due to auto-fluorescence in the wavelength range shorter than 475 nm
Solution Approach 1:
By changing the excitation wavelength parameter to 475 nm or longer through molecular structure modification, the patent shifts the operating wavelength away from the auto-fluorescence region (<475 nm). This parameter change maintains green fluorescence emission while avoiding excitation of background auto-fluorescence, thereby improving signal-to-background ratio
Solution Approach 2:
The invention replaces the conventional excitation wavelength approach (mechanical/optical parameter) with a chemically modified dye structure that intrinsically absorbs at longer wavelengths. This substitution fundamentally changes how green fluorescence is achieved, using molecular design rather than relying on short-wavelength excitation that triggers auto-fluorescence
3Measurement precision
If conventional sulfonated coumarin-based dyes are used, then fluorescence detection is enabled, but signal-to-background ratio is reduced due to high background fluorescence intensity
Solution Approach 1:
The patent changes the spectral parameters of the fluorescent dye by modifying its molecular structure (amino coumarin with sulfonate groups). This shifts the excitation maximum to 475-510 nm, creating a spectral separation between the dye's excitation and the auto-fluorescence region, thereby improving measurement precision and signal-to-background ratio in fluorescence detection
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 aminocoumarin compound effectively excites at longer wavelengths, enhancing fluorescence intensity and reducing background interference, thereby improving the signal-to-background ratio and enabling clearer, more accurate immunostaining results.
Implementation Method 1
The aminocoumarin compound is a fluorescent compound that emits light in a green region... the aminocoumarin compound is effectively excited at a wavelength of 475 nm or more... fluorescence spectrum maximum wavelength of 510 to 540 nm
Data Source
AI summary
The present invention provides: an aminocoumarin compound having a structure represented by the following formula (1); and an aminocoumarin compound-containing resin particle containing the aminocoumarin compound and a resin particle containing the aminocoumarin compound. In formula (1), Rs each independently represent a hydrogen atom or a methyl group, Q represents a sulfur atom, an oxygen atom, or N—R1, and R1 represents a hydrogen atom or a methyl group. The aminocoumarin compound of the present invention is a fluorescent dye that has a longer excitation wavelength than a conventionally known sulfonated coumarin-based compound and is effectively excited at a wavelength of 475 nm or more, particularly at a wavelength around 500 nm. The aminocoumarin compound of the present invention is a fluorescent dye that is excited, for example, at a wavelength of 475 to 510 nm and can be observed in a green light emission region of 510 to 540 nm using the wavelength characteristic, and can be expected to be suitably utilized in immunostaining or the like.