Cell Membrane Protein Staining Without Baking for Accurate Detection
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Solution Overview
Problem
Existing immunohistochemical staining methods for cell membrane proteins lack accuracy and specificity, particularly in detecting proteins like CAPRIN-1 for cancer diagnosis, due to the variability in abundance ratios and types among individuals.
Innovation Solution
A method that eliminates the baking step and includes embedding medium removal, antigen retrieval, primary and secondary antibody reactions, and label detection processes, utilizing a removal solution and temperature control to enhance staining accuracy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of substance
If baking is performed to remove embedding medium, then embedding medium removal is achieved, but cell membrane protein detection accuracy deteriorates
Solution Approach 1:
The invention changes the temperature parameter from high-temperature baking (typically 60-100°C) to low-temperature processing (room temperature or 4°C). This parameter change allows embedding medium removal while preserving cell membrane protein integrity and preventing denaturation, thereby resolving the contradiction between effective embedding medium removal and maintaining detection accuracy.
2Productivity
If high temperature is applied to remove embedding medium, then embedding medium removal efficiency is improved, but cell membrane protein structure is damaged
Solution Approach 1:
The invention introduces an embedding medium removal solution as an intermediary substance that facilitates embedding medium removal at low temperatures. This intermediary agent enables efficient removal without requiring high temperature, thus protecting cell membrane protein structure while achieving the removal goal.
Solution Approach 2:
The invention changes the temperature parameter from high-temperature processing to low-temperature processing (room temperature or 4°C). This parameter change maintains protein structure integrity while achieving effective embedding medium removal through the use of removal solutions, resolving the contradiction between removal efficiency and structural preservation.
3Adaptability or versatility
If conventional immunohistochemical staining is used, then general protein detection is achieved, but cell membrane protein specific detection is insufficient
Solution Approach 1:
The invention applies local quality by using low-temperature processing conditions specifically tailored for cell membrane protein preservation, while maintaining general immunohistochemical staining capabilities for other proteins. This localized optimization of temperature parameters enables specific cell membrane protein detection without sacrificing overall detection versatility.
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 achieves high specificity and accuracy in detecting cell membrane proteins, enabling precise determination of CAPRIN-1 expression for personalized cancer treatment.
Implementation Method 1
an embedding medium removal process of removing an embedding medium by contacting the fixed tissue comprising the embedding medium in a solid state with a removal solution
Implementation Method 2
a primary antibody reaction step of reacting a primary antibody having immunological reactivity against the cell membrane protein with the cell membrane protein of the fixed tissue
Implementation Method 3
a secondary antibody reaction step of reacting a labeled secondary antibody having immunological reactivity against the primary antibody with the fixed tissue after the primary antibody reaction step
Data Source
Figure 1A~1B
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Figure 3A~3B
AI summary
The present invention addresses the problem of providing a method for staining a cell membrane protein with high accuracy. Provided is a method for detecting a cell membrane protein in an embedded fixed tissue, said method comprising an embedded medium removal step for bringing a fixed tissue including a solid embedded medium into contact with a removal solution without baking the fixed tissue, thereby removing the embedded medium.