Modified Rhodamine Dyes for Multiplex Assays Using More Than Six Dyes
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Solution Overview
Problem
Existing rhodamine dyes have limited spectral characteristics, hindering the development of robust and sensitive multiplex assay systems with more than 6-dyes in combination.
Innovation Solution
Development of new rhodamine dyes with unique spectral properties, allowing for the creation of alternative multiplex dye sets by incorporating a class of compounds with specific structural modifications, including N-protected NH-rhodamine moieties and phosphate ester precursor groups, for labeling oligonucleotides.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If existing rhodamine dyes are used in multiplex assay systems, then the assays can be performed with current dye sets, but the spectral characteristics limit the ability to develop robust and sensitive assay systems using greater than 6-dyes in combination
Solution Approach 1:
The patent applies parameter changes by modifying the spectral characteristics of rhodamine dyes through structural modifications (N-protected NH-rhodamine moieties with specific substituents at positions 3, 6, 7, and 7'). These structural changes alter the emission and excitation spectra, enabling better spectral separation and allowing more dyes to be used in combination while maintaining assay robustness and sensitivity
Solution Approach 2:
The patent creates composite fluorescent dye sets by combining N-protected NH-rhodamine dyes with other fluorescent dyes in multiplex systems. The new rhodamine dyes serve as complementary components that fill spectral gaps, allowing the construction of composite dye sets with greater than 6 dyes that work together to enable high-plex multiplex assays
2Reliability
If new rhodamine dyes with unique spectral properties are developed, then more robust and sensitive multiplex assay systems can be created, but the complexity of dye set design and selection increases
Solution Approach 1:
The patent segments the fluorescent dye spectrum into distinct regions by developing rhodamine dyes with specific spectral profiles. Each N-protected NH-rhodamine dye is designed to occupy a particular spectral niche, allowing systematic selection and combination of dyes based on their spectral characteristics rather than trial-and-error approaches
Solution Approach 2:
The patent creates universal N-protected NH-rhodamine dye building blocks that can serve multiple functions in different multiplex assay configurations. The core rhodamine structure with N-protection provides a reliable spectral foundation that can be combined with various other dyes and used across different assay types, reducing the need for assay-specific dye optimization
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
Enables the creation of more robust and sensitive multiplex assay systems capable of using a higher number of dyes, enhancing the accuracy and efficiency of molecular biology and genetics assays.
Implementation Method 1
Fluorescent compounds are described that can be used to be label synthetic oligonucleotides
Data Source
AI summary
The present invention is directed to fluorescent rhodamine dyes having spectral properties suited to the creation of multiplex assay systems for use in molecular biology, cell biology and molecular genetics. The rhodamine dyes have the following structure.