Phase-Change Cell Immobilization for Loss-Free Staining
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Solution Overview
Problem
Conventional cell staining methods result in significant cell loss, particularly for rare cell types like circulating tumor cells, due to the lack of effective immobilization techniques, leading to difficulties in analysis and diagnosis.
Innovation Solution
A method and apparatus utilizing a phase change material to immobilize cells on a chamber surface by solidification, preventing cell loss during staining and washing, involving a chamber unit, bezel unit, and thickness adjustment unit to stabilize and control the gel thickness.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional pretreatment methods (cytospin or smear) are used to localize cells on slide surface, then cells can be positioned for staining, but 30% or more of cells are lost during staining reagent treatment and washing
Solution Approach 1:
A hydrogel layer is introduced as an intermediary between the slide surface and the cells. The hydrogel provides a protective matrix that anchors cells during staining and washing procedures, preventing cell loss while allowing reagent penetration. This mediator layer resolves the contradiction by enabling both cell positioning and high retention rates.
Solution Approach 2:
The physical state of the hydrogel is changed from liquid to gel through phase transition (temperature change or chemical crosslinking). This parameter change allows the hydrogel to transition from a fluid that can be easily applied to a solid-like structure that provides mechanical support and cell anchoring, thereby preventing cell loss during processing.
2Reliability
If plasma treatment is used to immobilize cells on membrane surface, then cell loss may be reduced, but equipment cost increases and reproducibility becomes remarkably low depending on cell type and charge value
Solution Approach 1:
The invention uses a simple, inexpensive hydrogel formulation that can be applied without complex plasma treatment equipment. The hydrogel serves as a disposable, easy-to-apply material that provides reliable cell immobilization through its gel matrix structure rather than electrostatic charges, eliminating the need for expensive plasma generators and reducing variability.
Solution Approach 2:
The hydrogel undergoes phase transition from liquid to gel state, providing a physical matrix for cell embedding. This phase change mechanism replaces the plasma treatment approach by using physical entrapment in the gel network rather than electrostatic adhesion, achieving reliable immobilization without complex equipment and with consistent results across different cell types.
3Loss of substance
If cells are immobilized in hydrogel by solidifying the phase change material, then cell loss during staining and washing is prevented, but additional steps and materials are required
Solution Approach 1:
The hydrogel application and cell immobilization steps are merged into a single process. The hydrogel is applied to the slide surface and simultaneously provides both the embedding matrix and the immobilization mechanism through its gel structure. This merging eliminates the need for separate immobilization steps and reduces the overall number of materials required.
Solution Approach 2:
The hydrogel serves multiple functions simultaneously: it acts as a mounting medium, an immobilization matrix, a protective layer during staining, and a washing support structure. This multi-functionality reduces the need for multiple separate materials and steps, achieving cell loss prevention without proportionally increasing complexity.
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 cell retention rates, facilitating precise analysis and isolation of single cells, especially for rare cell types, by minimizing cell loss during the staining process.
Implementation Method 1
immobilizing cells to be analyzed in a phase change material before staining
Implementation Method 2
solidifying the phase change material
Data Source
AI summary
Provided is a method and apparatus for cell staining without cell loss, and more particularly, to a method and apparatus for cell staining without cell loss during treatment of a staining reagent or washing reagent by immobilizing cells to be analyzed in a phase change material before staining. A method for cell staining without cell loss may effectively prevent cell loss during staining and analysis of rare cells that are becoming important in clinical diagnosis, and thus may facilitate observation, analysis and diagnosis thereby even with a trace amount of sample. In addition, as the method for cell staining without cell loss allows immobilization of cells, it makes discrimination and isolation of single cells easy, and may be effectively used in analysis of the isolated cells.