Cationic Borate Photoactivated Dye Bleaching
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Solution Overview
Problem
Current methods for analyzing biological samples are limited in their ability to detect multiple targets simultaneously due to the inefficiency of anionic borates in bleaching xanthene dyes and their susceptibility to oxidation, requiring special storage and handling protocols.
Innovation Solution
The use of a borate compound acting as an electron transfer agent, which undergoes a photoreaction upon irradiation to inactivate fluorescent dyes, allowing for the reuse of signal generators in subsequent cycles for detecting multiple markers in a biological sample.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If anionic borates with cationic counter ion are used for photoactivated chemical bleaching, then bleaching performance is improved, but susceptibility to oxidation increases and special storage protocols are required
Solution Approach 1:
The patent changes the chemical parameters of the borate compound by introducing positively charged substituents (such as quaternary ammonium groups) to the borate core structure. This parameter modification transforms the borate from anionic to cationic character, fundamentally altering its chemical properties including oxidation resistance while maintaining photoactivated bleaching capability
2Productivity
If anionic borates with cationic counter ion are used for iterative sample analysis, then signal inactivation is achieved, but storage and handling complexity increases
Solution Approach 1:
By changing the charge parameter of the borate compound to positive, the patent eliminates the need for special storage protocols associated with oxidizing agents. The cationic borates are stable under常规 storage conditions, simplifying the overall system complexity while maintaining the ability to perform iterative sample analysis
3Adaptability or versatility
If conventional borates are used for xanthene dye bleaching, then general applicability is maintained, but bleaching efficiency for xanthene dyes is insufficient
Solution Approach 1:
The patent applies local quality by designing borate compounds with specific positively charged functional groups (such as quaternary ammonium groups) positioned to interact optimally with xanthene dyes. This localized structural modification enhances bleaching efficiency for xanthene dyes specifically, while the core borate structure maintains general applicability to other fluorescent probes
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
This method enables high-throughput multiplex sample analysis by efficiently bleaching fluorescent dyes, allowing for the sequential analysis of multiple targets in a single sample without significant sample modification, improving the detection of presence, absence, concentration, and spatial distribution of biological targets.
Implementation Method 1
The PICB step can include applying a borate compound of Formula I, which acts as an electron transfer agent, and initiating a photoreaction, e.g., by irradiating the sample with visible light, to inactivate the signal generator
Implementation Method 2
initiating a photoreaction, e.g., by irradiating the sample with visible light, to inactivate the signal generator
Data Source
AI summary
Methods comprising the use of photoactivated chemical bleaching for detecting multiple targets in a biological sample are provided. The methods include the steps of providing a biological sample containing multiple targets, binding at least one probe to one or more target present in the sample, and observing a signal from the probe. The method further includes the steps of contacting the sample comprising the bound probe with a cationic or zwitterionic borate compound and irradiating the sample, thereby initiating a photoreaction that substantially inactivates the probe by photoactivated chemical bleaching. The method further includes the steps of binding at least one probe to one or more target present in the sample, and observing a signal from the probe. The process of binding, observing and bleaching may be iteratively repeated.