Phosphor-Tetrazine Monochromophore Probe for Multiplex Imaging
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Solution Overview
Problem
Conventional tetrazine-based fluorogenic probes experience significant reduction in fluorescence amplification efficiency in the long wavelength range (>600 nm), limiting their effectiveness for multiplex imaging due to energy transfer quenching issues.
Innovation Solution
A novel fluorogenic bioorthogonal probe with a phosphor-tetrazine monochromophore type, where the phosphor and tetrazine quencher share π-electrons as a single molecule, preventing energy transfer and maintaining high fluorescence amplification efficiency across all wavelength ranges.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional tetrazine-based fluorogenic probes use energy transfer quenching method, then fluorescence amplification efficiency is improved in short wavelength range, but fluorescence amplification efficiency significantly decreases in long wavelength range (>600 nm)
Solution Approach 1:
The patent changes the fundamental parameter of the probe structure from bichromophore (separated fluorophore and quencher) to monochromophore (integrated single molecule), which alters the energy transfer mechanism and enables high fluorescence amplification efficiency across all wavelength ranges including long wavelength (>600 nm)
Solution Approach 2:
Instead of using the conventional approach where fluorophore and quencher are electronically separated and connected via energy transfer, the patent inverts the design by integrating them into a single monochromophore molecule where the tetrazine unit is directly incorporated into the fluorophore structure, eliminating the need for energy transfer
2Ease of manufacture
If conventional probes use bichromophore type structure with separated phosphor and tetrazine, then synthesis is simplified, but energy transfer quenching occurs reducing fluorescence amplification efficiency
Solution Approach 1:
The patent merges the previously separated fluorophore and tetrazine quencher units into a single integrated monochromophore molecule, where the tetrazine unit is directly incorporated into the fluorophore structure. This combination eliminates energy transfer quenching while maintaining synthetic feasibility through modular construction of the integrated molecule
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 novel probe achieves high fluorescence signal amplification in all wavelength ranges, enabling effective multiplex imaging and specific labeling of targets, such as proteins, without the limitations of conventional bichromophore types.
Implementation Method 1
the previously developed fluorogenic bioorthogonal probe electronically separates the fluorescent molecule from the tetrazine, and then induced the quenching of the probe through an energy transfer method in which the excited energy of the fluorescent molecule is transferred to the tetrazine molecule
Implementation Method 2
a fluorogenic probe bound to tetrazine has been actively studied because it has rapid reactivity and high fluorescence amplification efficiency
Data Source
AI summary
The present invention relates to a novel phosphor-tetrazine and a use thereof and, more particularly, provides a novel compound having high fluorescence amplification efficiency in various wavelength ranges by using a compound having a novel core skeleton called tert-butyl (3-(7-(6-methyl-1,2,4,5-tetrazin-3-yl)-3-oxo-9-phenyl-1H-pyrrolo[3,4-b]indolizin-2(3H)-yl)propyl)carbamate.