Cyclic Fluorescence Imaging With Rapid De-Labeling Control
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Solution Overview
Problem
Existing methods for labeling and de-labeling molecules in biological contexts are inefficient, taking excessive time and introducing variability, especially when multiple cycles of labeling and de-labeling are required.
Innovation Solution
The use of ultrafast labeling and de-labeling methods involving controlled temperature, voltage application, and chemical quenching to rapidly label and remove detectable agents from cells or molecules, with systems incorporating temperature control, voltage sources, and detectors for efficient detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of time
If conventional labeling and de-labeling methods are used, then molecules can be labeled and detected, but the process takes excessive time and introduces variability
Solution Approach 1:
The patent applies parameter changes by optimizing temperature control (heating to 37°C or 60°C for specific durations), voltage application (1-100V for de-labeling), and chemical concentrations (detectable agent concentration, quenching agent concentration) to achieve rapid labeling and de-labeling within 15 minutes while maintaining consistency across cycles, thereby reducing both time loss and experimental variability
Solution Approach 2:
The patent implements periodic action through cyclic labeling and de-labeling procedures where cells are repeatedly labeled with detectable agents, imaged, then de-labeled and re-labeled with different agents. This systematic periodic process enables multi-plex detection while maintaining temporal control over each cycle to minimize variability
2Productivity
If multiple cycles of labeling and de-labeling are performed, then more molecules can be detected, but residual fluorescence increases and reduces detection accuracy
Solution Approach 1:
The patent converts the harmful effect of residual fluorescence into a benefit by implementing active quenching procedures. After imaging, cells are treated with quenching agents or subjected to voltage application that specifically removes or deactivates remaining detectable agents. This transforms the problem of residual signal into a controlled process that actively eliminates interference, enabling accurate sequential detection of multiple molecules
Solution Approach 2:
The patent employs chemical quenching agents that act as strong oxidants or reactive species generators to rapidly degrade or deactivate residual detectable agents on cell surfaces. This accelerated chemical removal process ensures complete elimination of background fluorescence between imaging cycles, maintaining high measurement precision while enabling detection of multiple target molecules
3Measurement precision
If detectable agents are applied to label cells, then molecules of interest can be identified, but de-labeling is inefficient and time-consuming
Solution Approach 1:
The patent replaces conventional mechanical or chemical washing methods for de-labeling with voltage-based removal. By applying electric fields (1-100V) across the cell layer, detectable agents are rapidly removed from cell surfaces through electrophoretic or electrostatic mechanisms. This substitution reduces de-labeling time from hours to minutes while maintaining complete removal efficiency, preserving both measurement precision and reducing time loss
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 rapid and efficient labeling and de-labeling of cells or molecules, reducing cycle times to under 15 minutes and minimizing residual fluorescence, suitable for high-throughput detection of multiple molecules with minimal experimental variability.
Implementation Method 1
applying a voltage across the cell or molecule; and de-labeling the cell or molecule, wherein de-labeling comprises removal of or quenching of the detectable agent on the cell or molecule
Implementation Method 2
heating the cell or molecule to a controlled temperature
Implementation Method 3
a detector configured to detect a first detectable agent
Data Source
AI summary
Methods and systems for improved labeling and/or de-labeling a molecule or cell in the context of scientific experimentation, industrial applications, and clinical investigation, including the means to repeat the process of labeling and de-labeling in an efficient manner.