One-Step Cell Staining Method Using Immobilizer and Permeabilization
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Solution Overview
Problem
Conventional cell staining methods are complex, leading to cell damage and loss, especially when detecting rare cells in liquid samples, and fail to efficiently stain specific substances expressed on cells.
Innovation Solution
A method where immobilization, permeabilization, and labeling treatments are performed in one step without washing, reducing the frequency of centrifugation and washing, and incorporating a sample collection tube design that separates immobilizer, permeabilization agent, and labeling substances to prevent cell damage and loss.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional multi-step staining procedures are used, then cells can be stained, but the procedure is complex and cells may be damaged or lost
Solution Approach 1:
The patent combines multiple separate treatment steps (immobilization, permeabilization, and labeling) into a single integrated step by co-incubating the cells with immobilizer, permeabilization agent, and labeled antibodies simultaneously in one buffer solution, eliminating the need for sequential processing and reducing cell handling operations
Solution Approach 2:
The patent uses a universal buffer solution that simultaneously performs multiple functions: maintaining cell structure during immobilization, creating permeability for antibody access, and providing the medium for antigen-antibody labeling reactions, thereby replacing multiple specialized solutions with one multi-functional reagent system
2Reliability
If multiple centrifugation and washing steps are performed, then cells can be processed, but rare cells may be damaged or flow out
Solution Approach 1:
The patent merges the functions of immobilization, permeabilization, and labeling into a single incubation step that requires no intermediate centrifugation or washing, eliminating cell loss associated with repeated pellet resuspension while maintaining complete processing functionality
Solution Approach 2:
The patent performs all necessary treatments (immobilization, permeabilization, and labeling) in a single pre-prepared buffer solution before any cell separation or processing steps, ensuring cells remain in solution throughout and are only pelleted once at the end for final observation
3Measurement precision
If conventional staining methods are used, then cells can be stained, but the time required is long
Solution Approach 1:
The patent combines immobilization, permeabilization, and labeling reactions into a single simultaneous incubation process that achieves complete staining in one step, reducing total procedure time while maintaining the specificity of antigen-antibody recognition through properly formulated buffer conditions
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 approach simplifies the cell staining process, reduces cell damage and loss, improves examination accuracy, and allows for efficient specific substance staining, while enabling simultaneous anticoagulation in blood samples.
Implementation Method 1
performing an immobilization treatment of the cells
Implementation Method 2
performing a permeabilization treatment of the cells
Implementation Method 3
reacted with an antibody
Data Source
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AI summary
An object of the present invention is, in the case when a specific substance that has expressed on cells to be detected in a liquid sample is to be specifically stained, to provide a convenient cell staining method by which the damage and loss (flow out) of the cells in the collected sample are prevented, and the time has been shortened in a procedure for staining the cells in the liquid sample. Furthermore, another object of the present invention is to provide a sample collection tube that is suitable for use in the above-mentioned cell staining method. A method for specifically staining a specific substance possessed by cells in a sample, including performing the following steps in one step: (A) a step of performing an immobilization treatment of the cells; and (B) a step of performing a permeabilization treatment of the cells.