Nanostructure FRET Labels for High-Plex Rare Cell Identification
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current multiplexing techniques in fluorescence microscopy and cytometry are limited by low plexing levels, subjectivity, and inter-operator variability, making it difficult to identify and isolate rare cells in biological samples effectively.
Innovation Solution
A label comprising a nanostructure backbone with attached dyes configured for energy transfer, allowing differentiation based on emission wavelength and fluorescence lifetime, enhancing the number of discernible labels through FRET pairs and diverse optical properties.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If traditional fluorescent dyes are used for marking target molecules, then the marking function is achieved, but the plexing level is limited to low numbers (1-5 for channel-based, 5-12 for spectral detectors)
Solution Approach 1:
The patent changes the parameters of the labeling system by using FRET pairs with different energy transfer efficiencies and fluorescence lifetimes. Instead of relying solely on spectral separation, the invention utilizes dynamic fluorescence parameters (lifetime, intensity ratios) that can be distinguished by time-resolved detection, thereby increasing the number of distinguishable labels beyond traditional spectral plexing limits
Solution Approach 2:
The patent introduces an intermediary mechanism (FRET energy transfer) between the excitation light and the emitted fluorescence. By using donor-acceptor dye pairs where energy transfer occurs, the system creates intermediate states with distinct fluorescence lifetimes and intensity ratios, enabling multiplexing beyond direct spectral detection capabilities
2Quantity of substance
If fluorescent cell barcoding with hue encoding is used, then multiplexing is achieved, but subjectivity and inter-operator variability increase
Solution Approach 1:
The patent replaces the subjective visual hue-based encoding system with an objective time-resolved fluorescence measurement system. Instead of relying on operators to visually distinguish and interpret color hues, the invention uses automated detection of fluorescence lifetimes and intensity ratios, which can be precisely measured and quantified without human subjectivity
Solution Approach 2:
The patent enables the labeling system to self-identify through intrinsic fluorescence properties (lifetime, intensity ratio) that are automatically measured by the detection system. The labels carry their own identification information in the form of distinct fluorescence decay characteristics, eliminating the need for subjective interpretation of encoded hues
3Quantity of substance
If more fluorescent dyes are used to increase plexing level, then the number of discernible labels increases, but the complexity of the marker system increases
Solution Approach 1:
The patent segments the labeling system into modular FRET pairs, where each pair consists of a donor dye and an acceptor dye with specific energy transfer characteristics. This modular approach allows systematic combination of pairs to generate multiple distinguishable labels while maintaining a structured, manageable system architecture rather than using arbitrary combinations of many individual dyes
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 identification and isolation of rare cells by increasing the number of distinguishable labels, improving the precision and reliability of cell type analysis in biological samples.
Implementation Method 1
The first dye and the second dye are within a distance from each other that enables an energy transfer between the first dye and the second dye
Data Source
AI summary
A label for usage in a marker for marking a target molecule in a biological sample includes a nanostructure backbone, and at least one group of dyes comprising a first dye and a second dye attached to the nanostructure backbone. The first dye and the second dye are within a distance from each other that enables an energy transfer between the first dye and the second dye. One of the dyes of the group of dyes is configured to emit light upon the energy transfer.