Horizontal Antigen Retrieval Using High-Boiling Solvents
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Solution Overview
Problem
Current antigen retrieval methods for immunohistochemical staining are energy-intensive, difficult to automate, and prone to evaporation issues, with enzymatic methods being dependent on enzyme stability and fixation time, and traditional heat-induced antigen retrieval requiring long procedures and high energy consumption.
Innovation Solution
A horizontal target retrieval method using a buffered solution with a solvent having a boiling point higher than water, which reduces the amount of solution and energy needed, eliminates the need for covering the sample, and simplifies the apparatus by heating the solution and sample above 100°C without evaporation issues.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If traditional heat-induced antigen retrieval is used, then antigenicity is restored, but energy consumption increases and procedure time increases
Solution Approach 1:
The patent changes the boiling point parameter of the retrieval solution by using organic solvents (ethanol, isopropanol, acetone) instead of water. This allows heating to temperatures above 100°C without water evaporation, achieving effective antigen retrieval in shorter times with lower energy consumption while maintaining reliability.
Solution Approach 2:
The patent accelerates the antigen retrieval process by using organic solvents with lower boiling points than water, allowing rapid heating and treatment cycles. This rushes through the traditional lengthy heating process while maintaining effective antigenicity restoration.
2Reliability
If traditional heat-induced antigen retrieval is used, then antigenicity is restored, but procedure time increases
Solution Approach 1:
The patent changes the thermal parameters by using organic solvents that can be heated to higher temperatures without evaporation. This enables faster treatment cycles and reduces the overall procedure time while maintaining reliable antigenicity restoration.
Solution Approach 2:
The patent skips the lengthy cooling and evaporation control steps required in traditional water-based methods by using organic solvents with higher boiling points, thereby rushing through the procedure while maintaining effectiveness.
3Reliability
If aqueous buffer is used for heating, then antigen retrieval is effective, but evaporation occurs requiring covering
Solution Approach 1:
The patent changes the solvent parameter from water to organic solvents with higher boiling points. This eliminates evaporation losses during heating while maintaining antigen retrieval effectiveness, removing the need for covering.
Solution Approach 2:
The patent converts the potential harm of evaporation into a benefit by using organic solvents that naturally resist evaporation at working temperatures. This eliminates the need for covering while maintaining solution volume and effectiveness.
4Ease of operation
If proteolytic enzymes are used for antigen retrieval, then epitope access is improved, but enzyme stability varies with storage time
Solution Approach 1:
The patent substitutes the biochemical mechanism of enzyme-based retrieval with a physical/chemical mechanism using organic solvents and heat. This replaces the unreliable enzyme system with a more stable physical process that does not degrade over time.
Solution Approach 2:
The patent changes the retrieval mechanism from enzymatic to thermal/chemical using organic solvents. This parameter change eliminates the stability issues associated with enzyme storage while maintaining effective epitope access.
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 method reduces energy and reagent consumption, shortens procedure time, and simplifies the apparatus, making it more economical and efficient while minimizing human error and evaporation problems, allowing for automated and reliable antigen retrieval.
Implementation Method 1
heating the tissue or cell sample and the buffered target retrieval solution to a temperature above 100° C.
Implementation Method 2
The high temperature accelerates hydrolysis of the aldehyde-induced modifications and the aldehyde released is diluted in the water
Data Source
AI summary
The present invention relates to a method for enhancing immunoreactivity of a tissue or cell sample fixed in a fixing medium, a target retrieval composition and its use. The method comprises providing a carrier in a horizontal position, said carrier having thereon a tissue or cell sample, said tissue or cell sample being on top of the carrier; contacting substantially the tissue or cell sample side of the carrier with a buffered target retrieval solution, wherein the target retrieval solution remains otherwise exposed to the environment; heating the tissue or cell sample and the target retrieval solution to a temperature above 100° C. The invention furthermore concerns the automated immunohistochemical staining of said samples, in particular the reactivation of antigens masked by fixation.