Biological Sample Mounting via Redox Treatment for Stable Adhesion
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Solution Overview
Problem
Existing methods for mounting biological samples on surfaces, such as microscope slides, often result in samples sticking too strongly or not strongly enough, leading to loss during incubation and staining processes, and can interfere with downstream analyses due to non-directed cross-linking of biomolecules.
Innovation Solution
A method involving the application of a reducing agent followed by an oxidizing agent to treat biological samples, reducing disulfide bridges and oxidizing thiol groups, enhancing sample adhesion to the surface.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If tissue sections are mounted on microscope slides using conventional coating methods (polylysine, collagen, gelatin, fibronectin, laminin, vitronectin, PEG), then adhesion of tissue sections to the slide is improved, but the coating has limited half-life of mostly a couple of months
Solution Approach 1:
The patent changes the chemical parameters of the slide surface by applying a silane coating that forms a permanent cross-linked network. This transforms the temporary organic coatings (polylysine, collagen, gelatin, fibronectin, laminin, vitronectin, PEG) into a stable inorganic-based surface modification that maintains its properties indefinitely, thus resolving the contradiction between adhesion improvement and coating durability.
Solution Approach 2:
The patent creates a composite surface structure by combining silane-based inorganic material with the glass slide substrate. This composite silane-coated surface provides both the adhesion properties needed for tissue mounting and the permanent durability required for long-term storage, eliminating the need for monthly re-coating of conventional organic coatings.
2Reliability
If formaldehyde is used to increase adherance of tissue sections to glass slides, then adhesion is improved, but non-directed cross-linking of biomolecules occurs that may interfere with downstream analyses
Solution Approach 1:
The patent extracts and eliminates the harmful formaldehyde cross-linking agent from the mounting process. Instead of using formaldehyde to achieve adhesion, the invention employs a silane coating that provides permanent attachment without causing non-directed cross-linking of biomolecules, thus removing the harmful factor while preserving the beneficial adhesion effect.
Solution Approach 2:
The silane coating acts as an intermediary layer between the glass slide and the tissue section. This intermediary surface provides the necessary adhesion through its chemical properties without directly interacting with or cross-linking the biomolecules in the tissue, thereby preventing the harmful non-directed cross-linking effect of formaldehyde while maintaining reliable attachment.
3Reliability
If plasma treatment or deposition of inorganic material (silanes or titanium dioxide) is used to modify glass surface, then stickiness of tissue sections is enhanced, but the process complexity increases
Solution Approach 1:
The patent employs a simple, cost-effective silane coating solution that can be applied directly to the slide surface without requiring complex plasma treatment equipment. This disposable-like simple coating approach provides effective tissue attachment while avoiding the high complexity and cost of plasma treatment or specialized deposition equipment, thus resolving the contradiction between enhanced stickiness and process complexity.
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
Improves the success of mounting and analyzing biological samples by ensuring better adhesion to the surface, reducing the risk of loss during staining and enhancing the effectiveness of downstream analytical techniques like microscopy and spatial transcriptomics.
Implementation Method 1
contacting said biological sample on said surface with a reducing agent
Implementation Method 2
contacting said biological sample on said surface with an oxidizing agent
Data Source
AI summary
The technology provided herein relates to methods for of mounting/sticking a biological sample on a surface by contacting said biological sample on said surface with a reducing agent.