Activatable Fluorogen Biosensor for Selective Voltage Imaging
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Solution Overview
Problem
Current voltage-sensitive probes diffuse into all brain cells, leading to non-selective monitoring of cell types and high background fluorescence, which hinders the ability to specifically target and detect voltage signals in neurological tissues.
Innovation Solution
Development of biosensor compositions comprising a FRET pair with an environment-sensitive donor and an activatable acceptor fluorogen, where the acceptor exhibits a >100-fold fluorescence increase when bound to a specific activator, allowing for targeted and reduced background fluorescence imaging.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If voltage-sensitive probes are used to monitor all cells in brain tissue, then complete network activity can be observed, but selective monitoring of specific cell types becomes impossible and background fluorescence increases
Solution Approach 1:
The patent applies local quality by making the probe's detection capability cell-type-specific through genetic targeting. The biosensor is expressed only in specific cell types via cell-type-specific promoters, so that different regions (cell types) have different detection qualities. This allows selective monitoring of specific cell types while maintaining the ability to study network activity by choosing appropriate promoters for different experimental goals
Solution Approach 2:
The patent segments the detection function by dividing the probe population into different biosensor variants, each targeted to specific cell types through different genetic promoters. This segmentation allows independent monitoring of different cell type networks simultaneously, resolving the contradiction between selective monitoring and comprehensive network observation
2Measurement precision
If voltage-sensitive probes are applied to all brain tissue, then optical signals from all excitable cells are obtained, but background fluorescence increases and detection sensitivity decreases
Solution Approach 1:
The patent extracts the harmful background fluorescence by using a genetically targeted expression system. Only cells that express the biosensor generate fluorescence signals, while all other cells remain dark. This extraction of the signaling function to specific cell types eliminates background fluorescence from non-target cells, thereby improving detection sensitivity
Solution Approach 2:
The patent introduces a genetic promoter as an intermediary that controls which cells express the biosensor. This promoter acts as a selective gatekeeper, allowing only specific cell types to become fluorescent when voltage changes occur. This intermediary mechanism resolves the contradiction by mediating between the desire for high signal and the need to eliminate background fluorescence
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 selective monitoring of specific cell types by reducing background fluorescence and enhancing signal detection sensitivity, revolutionizing optical neurobiology by allowing for precise imaging of voltage signals.
Implementation Method 1
The biosensors comprise a FRET (Förster resonance energy transfer or fluorescence resonance energy transfer) pair linked by a linker. A first member of the FRET pair is an environment-sensitive donor group and the second member is an activatable acceptor fluorogen. The environment-sensitive donor responds to light (electro-magnetic radiation not necessarily in the human visual spectrum) of a suitable excitation spectrum by transferring resonance energy to the activatable acceptor fluorogen, causing the activatable acceptor fluorogen to fluoresce.
Implementation Method 2
The activatable acceptor fluorogen produces a fluorescence signal increase of >100-fold when it interacts non-covalently with the activator as compared to when no activator is present.
Data Source
AI summary
Biosensor comprising an activatable acceptor fluorogen linked via a linker to a donor which transfers energy to the fluorogen on detecting an analyte wherein the fluorogen component reacts and a 100 fold increase in intensity results when the fluorogen interacts non-covalently with an activator e.g. fluorogen activator peptide.