Multiplex Imaging Optics With Peroxy Acid Fluorescence Quenching
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Solution Overview
Problem
Current multiplexed antibody-based fluorescence labeling methods for tissue imaging face various drawbacks that hinder their practical utilization.
Innovation Solution
An optical system for multiplex image acquisition using a microscope with a water dipping objective lens, a microfluidic chamber with a fluorinated ethylene propylene film viewing window, and a motorized stage for precise sample positioning, combined with a method to inactivate fluorophores using peroxy acids for rapid fluorescence quenching.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If traditional multiplexed antibody-based fluorescence labeling methods are used, then tissue imaging can be performed, but the process is time-consuming and suffers from residual fluorescence interference
Solution Approach 1:
The patent changes the chemical parameter by using peroxy acids instead of traditional hydrogen peroxide for fluorescence quenching. This parameter change results in significantly faster reaction kinetics, enabling rapid fluorescence removal and thereby improving imaging productivity while reducing processing time.
Solution Approach 2:
The patent replaces the mechanical/manual process of sequential imaging and manual fluorescence removal with an automated chemical system. The peroxy acid-based quenching method provides automated, rapid fluorescence removal that can be integrated into the imaging workflow, eliminating time-consuming manual intervention.
2Reliability
If traditional fluorescence quenching methods are used, then fluorescence can be removed, but the process is slow and incomplete
Solution Approach 1:
The patent changes the chemical reagent parameter from hydrogen peroxide to peroxy acids, which have superior quenching kinetics. This parameter change simultaneously improves both the speed and completeness of fluorescence quenching, resolving the contradiction between quenching efficiency and speed.
Solution Approach 2:
The patent employs peroxy acids as strong oxidizing agents that rapidly degrade fluorophores through oxidation. This accelerated oxidation mechanism provides both high-speed and high-efficiency fluorescence quenching, overcoming the limitations of traditional mild oxidizing agents.
3Measurement precision
If high-resolution imaging is performed, then detailed tissue structure can be visualized, but interference from residual fluorescence reduces image quality
Solution Approach 1:
The patent converts the harmful residual fluorescence into a beneficial outcome by using peroxy acids to completely and rapidly quench the fluorescence. This transforms the problem of fluorescence interference into an opportunity for achieving superior image quality with enhanced measurement precision.
Solution Approach 2:
The patent extracts and removes the harmful fluorescence signal from the imaging system through efficient chemical quenching. By completely removing the interfering fluorescence while preserving the tissue structure, the patent enables high-resolution imaging without interference.
4Adaptability or versatility
If multiple imaging cycles are performed for multiplex imaging, then multiple targets can be detected, but accumulated processing time reduces overall efficiency
Solution Approach 1:
The patent enables continuous multiplex imaging cycles by using rapid peroxy acid quenching that eliminates idle time between imaging rounds. The fast quenching allows the next antibody labeling and imaging cycle to begin immediately, maintaining continuous productive action and reducing total processing time while detecting multiple targets.
Solution Approach 2:
The patent optimizes the periodic cycling of multiplex imaging by shortening the quenching phase duration. The rapid peroxy acid quenching reduces the time each imaging cycle takes, allowing more cycles to be completed in the same overall time period, thereby improving both versatility and efficiency.
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 efficient and rapid fluorescence quenching, allowing for high-resolution multiplex imaging with enhanced precision and reduced interference, thereby improving the understanding of tissue and organ biology.
Implementation Method 1
an upright water dipping objective lens or water immersion lens configured to receive the light
Implementation Method 2
the viewing window has approximately the same refractive index as the solution in which the water-dipping objective is immersed
Implementation Method 3
a method to inactivate fluorophores using peroxy acids for rapid fluorescence quenching
Data Source
AI summary
The disclosure features systems and methods for multiplex imaging. In particular, improved systems and methods for multiplex imaging which are faster and more efficient than those known in the art are disclosed herein.