Violet Laser Excitable Dyes with Extended Stokes Shift
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Solution Overview
Problem
Current violet laser-excitable dyes have weak absorptivity, low quantum yields, and tend to aggregate in aqueous solutions, leading to significant residual fluorescence background and decreased detection sensitivity in biological applications.
Innovation Solution
Development of novel dye compounds with a reactive group attached to a fluorescent nucleus, exhibiting high solubility and a long Stokes shift, allowing for efficient excitation in the 350-500 nm range and minimizing quenching, thereby enhancing detection sensitivity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Use of energy by moving object
If existing violet laser excitable dyes are used, then they can be excited by 405 nm laser, but they have weak absorptivity and low quantum yields
Solution Approach 1:
The patent modifies the chemical structure of the dye by changing parameters such as introducing electron-donating groups at specific positions on the xanthene ring system and adjusting the substituent groups (R1-R10) to optimize both absorptivity and quantum yield simultaneously
Solution Approach 2:
The invention creates composite fluorescent compounds by combining the xanthene core structure with various reactive groups and substituent molecules, achieving synergistic effects where the composite structure provides both high absorptivity and high quantum yield
2Use of energy by moving object
If existing violet laser excitable dyes are used, then they can be excited by 405 nm laser, but they aggregate in aqueous solutions
Solution Approach 1:
The patent introduces hydrophilic substituent groups at specific local positions on the dye molecule to improve water solubility without affecting the core fluorescent xanthene structure's excitation properties
Solution Approach 2:
The invention uses solubilizing groups as intermediary structures that bridge the hydrophobic xanthene core and the aqueous environment, enabling the dye to remain dissolved in water-based biological samples
3Use of energy by moving object
If existing violet laser excitable dyes are used, then they can be excited by 405 nm laser, but they produce significant residual fluorescence background
Solution Approach 1:
The patent modifies the emission spectrum of the dye by adjusting the xanthene ring substituents to shift the emission wavelength away from the violet laser excitation wavelength, creating a larger Stokes shift and reducing background fluorescence
4Use of energy by moving object
If existing violet laser excitable dyes are used, then they can be excited by 405 nm laser, but they have decreased detection sensitivity
Solution Approach 1:
The invention optimizes multiple parameters including molecular weight, substituent groups, and conjugation length to enhance the fluorescence signal intensity and improve detection sensitivity while maintaining 405 nm excitation capability
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 new dye compounds provide brighter signals with improved stability and water solubility, reducing background fluorescence and increasing detection sensitivity in multiplexed applications such as microscopy and flow cytometry.
Implementation Method 1
Fluorescent dyes are widely used as tracers for localization of biological structures by fluorescence microscopy... the detectability of emission at a wavelength distinct from the excitation, the orders of magnitude greater detectability of fluorescence emission over light absorption
Implementation Method 2
the absorptivity (extinction coefficients of less than 20,000 cm−1 M−1 at their absorbance maxima)... weak absorbtivity
Data Source
AI summary
The present invention provides dye compounds optimally excited at about 400 nm and have a Stokes shift of at least about 80 nm. These dyes find use in detection of analyte in a sample and the preparation of dye-conjugates.