Fluorescent Label Preservation Using Oil-Based Mounting Medium
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Solution Overview
Problem
Current methods for tissue staining using fluorescent labels face challenges with light resistance, particularly when using oil-based mounting media, which can lead to deterioration of the fluorescent label and impaired staining properties, necessitating the use of aqueous media for fluorescent staining.
Innovation Solution
The method involves immobilizing the specimen after fluorescent labeling and using an oil-based mounting medium with a discoloration inhibitor to improve light resistance and maintain staining properties, allowing for permanent preparations with good color tone and color development.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Duration of action of stationary object
If oil-based mounting medium is used for fluorescent staining, then permanent preparation with good color tone is achieved, but fluorescent label deteriorates and staining properties are impaired
Solution Approach 1:
A water-soluble polymer is introduced as an intermediary substance between the fluorescent label and the oil-based mounting medium. This polymer forms a protective barrier that prevents direct contact between the oil and fluorescent label, thereby preventing deterioration while allowing the oil-based medium to provide its benefits for permanent preparation and color tone.
Solution Approach 2:
The mounting system is transformed into a composite structure consisting of three components: the fluorescent label, the water-soluble polymer coating, and the oil-based mounting medium. This composite approach allows each component to perform its optimal function - the polymer protects the fluorescent label while the oil medium provides stability and optical properties.
2Reliability
If aqueous mounting medium is used for fluorescent staining, then fluorescent staining properties are maintained, but permanent preparation and good color tone are compromised
Solution Approach 1:
The water-soluble polymer acts as a mediator that enables the use of oil-based mounting medium while preserving fluorescent staining properties. By coating the fluorescent label with this polymer before mounting, the system achieves both fluorescent stability and permanent preparation capability.
3Illumination intensity
If high-intensity excitation light is used for fluorescence observation, then observation capability is improved, but fluorescent label light resistance decreases
Solution Approach 1:
The water-soluble polymer is applied beforehand to the fluorescent label to create a protective cushion or barrier. This pre-protection mechanism shields the fluorescent label from the damaging effects of high-intensity excitation light during observation, thereby extending its lifespan and maintaining light resistance.
Solution Approach 2:
The polymer coating serves as an intermediary layer between the fluorescent label and the excitation light, absorbing or scattering harmful light energy before it reaches the fluorescent label, thus protecting it from photodegradation.
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 enables the preparation of specimens with preserved fluorescent staining properties and improved light resistance, even when using oil-based mounting media, facilitating reliable fluorescence microscopy observations.
Implementation Method 1
irradiating the thus stained pathological specimen with excitation light to allow fluorescence to be emitted for detection of a biological substance
Implementation Method 2
mounting the thus immobilized specimen using an oil-based mounting medium comprising a discoloration inhibitor
Data Source
AI summary
The present invention provides a staining method in which the fluorescent staining properties in a fluorescently-immunostained specimen are not reduced even when an oil-based mounting medium is used. The present invention also provides a method of preventing deterioration of a fluorescent label caused by irradiation with excitation light and improving the light resistance in a fluorescently-immunostained specimen obtained by the staining method. The biological substance detection method according to the present invention is a biological substance detection method for specifically detecting a biological substance from a pathological specimen, which includes the steps of: immunostaining the specimen with a fluorescent label; immobilizing the thus stained specimen; and mounting the thus immobilized specimen using a mounting medium including an organic solvent not freely miscible with water. In the biological substance detection method, the above-described mounting medium further includes a discoloration inhibitor.