Fluorescent Labelling Device Segmented Reagent Reservoirs
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Solution Overview
Problem
The short functional life of OPA/BME labelling mixtures due to rapid oxidation of β-mercaptoethanol (BME) at high pH limits the duration of fluorescent labelling experiments, especially in remote locations, and requires manual addition of BME shortly before use, making it impractical.
Innovation Solution
A method involving separate strongly buffered alkaline and weakly buffered acidic solutions for ortho-dialdehyde dye and reducing agent, respectively, with optional antioxidant additives, stored in impermeable reservoirs and combined via a network of connection channels to optimize pH conditions and extend the functional life of the reducing agent.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If OPA/BME mixture is stored at high pH for labelling reactions, then labelling efficiency is improved, but the functional life of the reducing agent decreases rapidly due to oxidation
Solution Approach 1:
The invention divides the OPA/BME mixture into two separate reservoirs: one containing the alkaline OPA solution and another containing the acidic BME solution. This segmentation prevents premature mixing and oxidation of BME at high pH, allowing each component to be stored under optimal conditions independently before combination during use.
Solution Approach 2:
The invention changes the pH parameter of the BME solution from alkaline to acidic (low pH) during storage. This parameter change dramatically reduces the oxidation rate of BME, extending its functional life from approximately one week at high pH to several months or longer at low pH. During labelling, the pH is temporarily raised by mixing with alkaline OPA solution.
2Manufacturing precision
If manual addition of BME is required shortly before use, then labelling quality is maintained, but operational complexity and user burden increase
Solution Approach 1:
The device is designed to automatically mix the alkaline OPA solution and acidic BME solution through a network of connection channels when activated. This self-service mechanism eliminates the need for users to manually add BME shortly before use, while still maintaining optimal labelling quality through controlled mixing ratios and timing.
3Ease of operation
If OPA/BME mixture is prepared in advance for remote location experiments, then experimental convenience is improved, but the mixture becomes ineffective due to BME oxidation
Solution Approach 1:
By segmenting the mixture into separate reservoirs containing alkaline OPA and acidic BME solutions, the invention enables advance preparation and transport of stable reagent components to remote locations. The solutions remain effective for months during storage and transport, then combine reliably when needed to perform labelling experiments.
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 approach extends the functional life of the reducing agent by up to 75 days without additives and up to 500 days with antioxidants, enabling longer-term fluorescent labelling and reducing the need for immediate BME addition.
Implementation Method 1
reacting the component with the aromatic ortho-dialdehyde dye and the reducing agent
Implementation Method 2
fluorescently label the components... OPA yields fluorescently labelled biomolecules by reacting with primary amine groups
Implementation Method 3
atmospheric oxygen has been known to cause BME to oxidise rapidly
Implementation Method 4
reacting the component with the aromatic ortho-dialdehyde dye and the reducing agent
Data Source
AI summary
A device and method for fluorescent labelling a component is provided. The device comprising a first user-replaceable reservoir comprising a strongly buffered alkaline solution of aromatic ortho-dialdehyde dye, a second user-replaceable reservoir comprising a weakly buffered acidic solution of a reducing agent, one or more fluid pathways comprising the component, and a network of connection channels linking the reservoirs and the fluid pathway to enable the alkaline solution and the acidic solution to combine with the component in order to label the component by reacting the component with the alkaline solution and the acidic solution.