Antigen Activation Solution Using Urea for Cytodiagnosis
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Solution Overview
Problem
Conventional methods for activating antigens in liquid-phase fixed cells require heat, which can damage cells and hinder precise diagnosis in cytodiagnosis, especially when using proteolytic enzymes like trypsin, pepsin, or papain.
Innovation Solution
A nonbridging fixation solution containing agents that break hydrogen bonds, such as urea, is used to activate antigens without heat, maintaining cell morphology and allowing antigen-antibody combination, with a preferred concentration of urea between 10 to 15 w/v % and a pH range of 7 to 9, along with a buffer and chelator to facilitate antigen exposure.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If heat is applied to activate antigen in liquid-phase fixed cells, then antigen activation is achieved, but cell morphology is damaged
Solution Approach 1:
The invention changes the activation mechanism from thermal energy (heat) to chemical energy by introducing proteolytic enzymes (trypsin, pepsin, or papain) that specifically break hydrogen bonds at pH 7-9, enabling antigen activation without temperature increase and thus preserving cell morphology
Solution Approach 2:
The invention replaces the thermal mechanical system (heat application) with a biochemical system (proteolytic enzyme action), substituting physical energy input with chemical catalysis to achieve the same antigen exposure effect while avoiding thermal damage to cell structure
2Reliability
If proteolytic enzymes are used to activate antigen, then antigen exposure is improved, but cell damage occurs
Solution Approach 1:
The invention optimizes the pH parameter to the range of 7-9, which is physiological and compatible with cell structures, allowing proteolytic enzymes to function effectively at mild conditions that minimize cellular damage while achieving sufficient antigen exposure
Solution Approach 2:
The invention uses mild concentrations of proteolytic enzymes and controls treatment time to achieve partial digestion sufficient for antigen exposure without excessive enzymatic action that would damage cell morphology, applying just enough enzyme activity to break hydrogen bonds while preserving cellular integrity
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 effectively activates antigens without heat damage, improving the efficiency of immunostaining and maintaining cell morphology, allowing for precise cytodiagnosis by breaking hydrogen bonds and removing impurities, thus enhancing the detection of cells in liquid-based cytology.
Implementation Method 1
hydrogen bonds are present between antigen molecules, between an antigen molecule and nearby molecules, and/or between an antigen molecule and surrounding molecules in the liquid-phase fixed sample, and that cutting of these hydrogen bonds (application of energy exceeding the enthalpy of the hydrogen bonds) exposes the antigen molecules (activates the antigen)
Data Source
AI summary
A method of activating an antigen is herein described. The method comprises providing an antigen activation solution and contacting the cell with the solution to activate the antigen. A method of detecting a cell fixed by a nonbridging fixation solution is also described. The method comprises providing an antigen activation solution, contacting the cell with the solution, immunostaining the cell, and detecting the stained cell. The solution used for these methods comprises an agent for breaking a hydrogen bond, and is also described herein.