Fluorescent Staining Method for Glycol Groups
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Solution Overview
Problem
Current methods for fluorescent staining of substances containing glycol groups in tissues or cells, especially for structures like mucus or basement membranes, face challenges in achieving clear three-dimensional imaging due to low tissue permeability of excitation light, limiting their effectiveness in histopathological diagnosis and research.
Innovation Solution
A fluorescent staining method using a compound with a fluorescein skeleton, compatible with PAS staining, is developed, which combines with tissue clearing technologies to enable clear three-dimensional observation by optimizing the excitation wavelength for improved tissue permeability and imaging.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a fluorescent probe with short excitation wavelength is used for staining substances containing glycol groups, then the staining specificity is improved, but the tissue permeability of excitation light deteriorates, making clear three-dimensional imaging difficult
Solution Approach 1:
The patent changes the excitation wavelength parameter from short wavelength to long wavelength (blue or green excitation) to improve tissue permeability while maintaining staining specificity through the use of fluorophores with appropriate excitation spectra. This parameter change allows penetration of excitation light through deeper tissue layers for three-dimensional imaging.
Solution Approach 2:
The patent employs composite fluorescent probes combining specific chemical moieties (for glycol group binding) with fluorophores having long excitation wavelengths. This composite structure integrates the staining specificity function with the light penetration function, resolving the contradiction between specificity and tissue permeability.
2Ease of operation
If conventional fluorescent probes are used for whole-mount staining, then the staining procedure is simplified, but the ability to achieve clear three-dimensional observation deteriorates
Solution Approach 1:
The patent changes the optical parameters of the fluorescent probe (using long excitation wavelength fluorophores) to enable clear three-dimensional imaging while maintaining the simplicity of whole-mount staining procedures. The fluorophores are designed to be compatible with standard staining protocols while providing superior optical properties for 3D observation.
3Measurement precision
If existing fluorescent probes are combined with tissue clearing technology, then three-dimensional imaging is enabled, but the adaptability to clinical pathological diagnosis deteriorates due to lack of PAS staining compatibility
Solution Approach 1:
The patent designs fluorescent probes that serve multiple functions: they are compatible with both PAS staining procedures and tissue clearing technologies, while enabling three-dimensional imaging. This multi-functional probe design allows it to adapt to various diagnostic and research applications without requiring separate specialized probes for each technique.
Solution Approach 2:
The patent adjusts the chemical and optical parameters of the fluorescent probe to ensure compatibility with PAS staining chemistry while maintaining long excitation wavelength properties for 3D imaging. This parameter optimization allows the probe to work across multiple staining protocols and imaging requirements.
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
The method allows for specific and clear three-dimensional visualization of substances with glycol groups within tissues or cells, enhancing diagnostic accuracy and research capabilities in histopathology and life sciences.
Implementation Method 1
a compound having a fluorescein skeleton enables special staining compatible with PAS staining, and the combination thereof with a tissue clearing technology and a 3D imaging technology enables clear three-dimensional observation
Data Source
AI summary
A fluorescent staining method compatible with PAS staining that is special staining including staining a structure (e.g., mucus or a basement membrane) containing a large amount of a substance containing a glycol group such as a polysaccharide, the fluorescent staining method enabling clear three-dimensional observation. A method of fluorescently staining a substance containing a glycol group in a tissue or cell sample, the method including bringing a compound represented by the following general formula (I) or a salt thereof and the tissue or cell sample into contact with each other.