Evaporation Reducing Agent for Tissue Assay Reagent Stability
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Solution Overview
Problem
Immunohistochemical assays and in situ hybridization techniques face challenges with evaporation of assay reagents at elevated temperatures, particularly in small volumes, which can compromise sample integrity and assay accuracy.
Innovation Solution
A reagent containing an evaporation reducing agent, such as ethylene glycol or other compounds like glycine, is used in an aqueous solution to minimize evaporation by reducing the vapor pressure and boiling point of the solvent, thereby limiting reagent loss to 20% or less during processing steps like hybridization and antigen retrieval.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If elevated temperature is applied during assay processing, then hybridization and denaturation efficiency are improved, but reagent evaporation increases
Solution Approach 1:
The patent introduces an evaporation reducing agent as an intermediary substance that mediates between the thermal processing requirements and evaporation prevention needs. This agent is added to the aqueous assay reagent to form a modified reagent composition that maintains the necessary thermal properties for hybridization while suppressing evaporation through vapor pressure reduction.
Solution Approach 2:
The patent modifies the physical-chemical parameters of the assay reagent by adding evaporation reducing agents that change the vapor pressure and boiling point characteristics. This parameter modification allows the reagent to withstand elevated temperatures without excessive evaporation, thereby resolving the contradiction between thermal processing efficiency and reagent retention.
2Measurement precision
If small volumes of reagent are used, then assay precision is improved, but evaporation loss becomes more significant
Solution Approach 1:
The evaporation reducing agent serves as a protective intermediary that enables the use of small reagent volumes by compensating for the heightened evaporation risk. The agent modifies the reagent's evaporation characteristics so that even small volumes maintain sufficient quantity throughout the assay process, preserving both precision and adequacy.
Solution Approach 2:
The patent applies beforehand cushioning by pre-adds the evaporation reducing agent to the reagent before the assay begins. This proactive measure creates a protective effect that cushions against evaporation losses throughout the subsequent heating steps, ensuring that small volumes do not evaporate excessively and compromising assay precision.
3Duration of action of moving object
If longer processing time is used, then hybridization completeness is improved, but total evaporation loss increases
Solution Approach 1:
The evaporation reducing agent acts as a time-extending intermediary that allows longer processing durations without proportional increases in evaporation loss. By modifying the reagent's evaporation resistance, the agent enables extended hybridization times to achieve completeness while the evaporation rate remains suppressed throughout the extended period.
Solution Approach 2:
The patent ensures continuity of useful action by maintaining stable reagent volumes throughout extended processing times. The evaporation reducing agent provides continuous protection against evaporation, allowing the hybridization process to proceed continuously over longer periods without interruption or volume loss that would compromise completeness.
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 use of evaporation reducing agents effectively reduces reagent loss during elevated temperature steps in immunohistochemical assays, maintaining assay performance and sample integrity by limiting evaporation to 20% or less, ensuring consistent results across various temperatures and processing times.
Implementation Method 1
minimize evaporation by reducing the vapor pressure and boiling point of the solvent
Implementation Method 2
minimize evaporation by reducing the vapor pressure and boiling point of the solvent
Data Source
AI summary
A method including contacting a tissue or cellular sample with a reagent including an evaporation reducing agent(s) having the general formula X1—R—X2, wherein R is an alkyl, alkenyl, alkynyl or an aromatic moiety of 1 or more carbon atoms that may be substituted with an oxygen, nitrogen or sulfur and X1 and X2 are independently selected to be moiety that is susceptible to hydrogen bonding and processing the tissue or cellular sample. A reagent used in the processing of a tissue or cellular sample with the reagent containing an evaporation reducing agent.