DHP-Cyanine and Squaraine Dyes for Broad-Spectrum Flow Cytometry
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Solution Overview
Problem
Existing dihydrophenanthrene-based polymer dyes have strong excitation in the ultraviolet and violet regions, and attempts to shift excitation to the red region face synthetic challenges and retention of original absorption, limiting their applicability in multicolor panels for flow cytometry.
Innovation Solution
Development of DHP-cyanine and DHP-squaraine compounds and polymers with water solubility, capable of being excited across UV, violet, blue, yellow, green, red, and near-infrared wavelengths, achieved through modifications to the dihydrophenanthrene monomers and incorporation of fused heterocyclic rings, allowing for fine-tuning of absorption and emission.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If attempts are made to shift excitation to the red region by incorporating modifier unit monomers into existing DHP violet-excitable polymer backbones, then red region excitation is achieved, but the original UV and violet absorption is retained along with strong UV and violet emission from the core
Solution Approach 1:
The patent extracts the problematic core DHP structure that causes unwanted UV/violet emission and replaces it with modified monomers containing fused heterocyclic rings. This removes the harmful emission source while preserving the desired red region excitation capability through the modified monomer design.
Solution Approach 2:
The patent changes the chemical structure parameters of the monomers by incorporating fused heterocyclic rings (such as indole, quinoline, benzoxazole, benzothiazole) which fundamentally alter the absorption and emission characteristics. This structural parameter change enables excitation in the red region while eliminating unwanted UV/violet emission.
2Adaptability or versatility
If attempts are made to design red shifted core polymers using other aromatic cyclic molecules as monomers, then red shifted excitation is achieved, but synthetic challenges arise
Solution Approach 1:
The patent segments the monomer structure into a core DHP unit combined with specific fused heterocyclic ring systems. This segmentation allows for systematic design and synthesis where each component can be optimized independently, reducing overall synthetic complexity while achieving red-shifted excitation.
Solution Approach 2:
The patent creates composite monomer structures by combining DHP cores with fused heterocyclic rings (indole, quinoline, benzoxazole, benzothiazole). These composite structures provide both the desired red-shifted optical properties and improved synthetic accessibility compared to entirely new aromatic cyclic molecules.
3Adaptability or versatility
If more careful design is required for shifting excitation maxima to blue, green, and red regions, then broader spectral coverage is achieved, but device complexity and design requirements increase
Solution Approach 1:
The patent develops a universal platform of DHP-based monomers with fused heterocyclic rings that can be systematically modified to achieve different excitation wavelengths across blue, green, and red regions. This universal design approach reduces overall complexity by providing a consistent structural framework that can be adapted for multiple spectral regions rather than requiring entirely separate dye designs for each region.
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 DHP-cyanine and DHP-squaraine compounds and polymers provide a range of excitation maxima from 400-900 nm, enabling their use in flow cytometers and spectral flow instruments, enhancing multicolor detection capabilities.
Implementation Method 1
The DHP-Cy and DHP-squaraine compounds, polymers, labeled specific binding partners, and tandem dyes according to the present disclosure can be excited using UV, violet, blue, yellow, green, red, or NIR wavelengths
Implementation Method 2
Water soluble fluorescent compounds and their conjugates can be used in a variety of biological applications by generating signals which can be monitored in real time
Data Source
AI summary
The present disclosure provides novel dihydrophenanthrene (DHP)-cyanine and DHP-squaraine fluorescent compounds and water-soluble polymers thereof. The DHP-cyanine and DHP-squaraine fluorescent compounds and polymers can be excited using UV, violet, blue, yellow, green, red, or NIR wavelengths. The fluorescent dyes may be conjugated to antibodies for detection of target analytes in biological samples and are suitable for use in flow cytometry analyses.