Peptide-Backbone Fluorescent Labels That Prevent Self-Quenching
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Solution Overview
Problem
Developing small fluorescent labels with high brightness is challenging due to self-quenching issues arising from dye-dye interactions, which affect the quality of fluorescence imaging, especially in super-resolution and 3D microscopy.
Innovation Solution
Designing fluorescent probes with a peptide backbone that maintains a sufficient distance between fluorescent moieties, using a general formula Y-(L-(A(F)-Pi) n -A(F)-P') m, where Pi comprises 10-30 amino acids, to minimize dye-dye interactions and prevent self-quenching.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Illumination intensity
If conventional fluorescent labels are used, then brightness is improved, but self-quenching occurs due to dye-dye interactions
Solution Approach 1:
The fluorescent label is segmented into distinct components: a peptide backbone (Y) and separately positioned fluorescent moieties (F) connected via linkers (L). This segmentation ensures that fluorescent dyes are spatially separated, preventing dye-dye interactions and self-quenching while maintaining high brightness through proper positioning.
Solution Approach 2:
The peptide backbone acts as an intermediary structure that connects fluorescent moieties at controlled distances. The linker units (L) serve as mediators between the peptide backbone and fluorescent dyes, ensuring optimal spacing that prevents self-quenching while allowing sufficient fluorescence signal for high-quality imaging.
2Length of moving object
If small fluorescent labels are used, then tissue penetration is improved, but brightness may be reduced
Solution Approach 1:
The invention changes the structural parameters of fluorescent labels by using peptide backbones with controlled lengths (10-30 amino acids) and optimizing the number and positioning of fluorescent moieties. This parameter optimization achieves a balance between small size for tissue penetration and sufficient brightness for high-quality imaging through multiple strategically positioned fluorophores.
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 probes provide specific staining and enhanced tissue penetration, resulting in higher brightness and faster imaging speeds, particularly in light-sheet and super-resolution microscopy, with reduced self-quenching effects.
Implementation Method 1
Fluorescence microscopy is a powerful technique used to generate high-content imaging datasets
Data Source
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AI summary
The invention is directed to fluorescent probes comprising an antigen recognizing moiety covalently bound by a peptide backbone to at least one fluorescent dye characterized in that the fluorescent probes have the general formula I Y-(L-(A(F)-Pi)n-A(F)-P')m With Pi: same or different peptide comprising 10 - 30 amino acids, P': a peptide comprising 0 - 50 amino acids F: fluorescent moiety A: an amino acid, L: a linker unit Y: antigen or antigen recognizing moiety m: 1-100