Fluorescent Compounds Large Stokes Shift Cellular Imaging

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Solution Overview

Problem

Current fluorescent probes for cellular and organelle imaging have limitations such as low sensitivity, cytotoxicity, high cost, and inability to detect environmental polarity changes, with small Stokes shifts causing spectral overlap and interference in biological applications.

Innovation Solution

Development of a family of fluorophores with large Stokes shifts, high environmental stability, and solvatochromic properties, allowing for selective cellular and subcellular labeling, imaging, and analysis, particularly suitable for diagnosing chronic stress and lipid-related metabolic disorders, which are highly membrane-permeant, non-toxic, and easily synthesized.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If fluorescent probes with small Stokes shifts are used, then the excitation and emission wavelengths are close, but this causes spectral overlap and interference in biological applications

Engineering Contradiction:
Improvedetection precisionVSAvoidspectral interference
Core Design Contradiction:
Measurement precisionVSObject-generated harmful factors

Solution Approach 1:

The patent applies parameter changes by designing fluorophores with significantly increased Stokes shift values (changing the wavelength difference parameter between excitation and emission). This parameter modification separates the excitation and emission spectra, eliminating spectral overlap and interference while maintaining detection precision.

Inventive Principle:
Principle #35Parameter changes

2Reliability

If conventional fluorescent probes are used for cellular imaging, then labeling can be achieved, but cytotoxicity and low environmental stability limit their application

Engineering Contradiction:
Improveenvironmental stabilityVSAvoidcytotoxicity
Core Design Contradiction:
ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent employs composite materials by creating fluorophore molecules with specific structural compositions (including heterocyclic rings, conjugated systems, and stabilizing substituents) that simultaneously provide environmental stability and reduced cytotoxicity. The composite molecular structure integrates multiple functional elements that work together to enhance reliability while minimizing harmful effects on biological systems.

Inventive Principle:
Principle #40Composite materials

3Measurement precision

If fluorescent probes are used to detect biological molecules, then sensitivity is improved, but the cost and complexity of probe synthesis increase

Engineering Contradiction:
Improvedetection sensitivityVSAvoidsynthesis complexity
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The patent applies segmentation by dividing the fluorophore synthesis into modular stages with standardized intermediate compounds. The modular molecular structure allows for systematic assembly of fluorophores with desired properties, reducing synthesis complexity while maintaining detection sensitivity. Common intermediates can be reused across different fluorophore preparations.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The patent utilizes parameter changes by optimizing synthesis conditions (temperature, catalysts, reaction time) to achieve high sensitivity fluorophores through simplified reaction pathways. By changing key synthesis parameters, the patent reduces the number of steps required while maintaining or enhancing detection sensitivity.

Inventive Principle:
Principle #35Parameter changes

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

These fluorophores provide clear distinction between excitation and emission wavelengths, reducing interference and enhancing signal strength, enabling effective visualization and analysis of cellular structures and lipid compositions with low cytotoxicity and cost-effectiveness.

Implementation Method 1

Fluorescence microscopy and fluorescence imaging are expanding areas of research

Methodology Applied
Scientific EffectFluorescence: Fluorescence

Implementation Method 2

The Stokes shift is the wavelength distance between the excitation and emission wavelengths of a given compound

Methodology Applied
Scientific EffectStokes shift:

Implementation Method 3

The compounds now disclosed also present solvatochromism, depending on the solvent polarity

Methodology Applied
Scientific EffectSolvatochromism:

Implementation Method 4

a family of fluorophores, which permeates cell membranes enabling the labelling of fixed or living cells

Methodology Applied
Scientific EffectPermeation: Permeation

Data Source

PatentEP3445348B1Fluorescent compounds, production methods and uses thereof
Publication Date: 2023.06.07 UNIV AVEIRO
  • EP3445348B1 patent drawingFigure 1~2
  • EP3445348B1 patent drawingFigure 3~4
  • EP3445348B1 patent drawingFigure 5~6B

AI summary

The present disclosure relates to fluorescent compounds with large Stokes' shift. This disclosure also relates to the, production methods and use of said fluorescent compounds for cellular labelling and in vivo imaging for use in medicine or for use in cell culture.