Phase-Change Cell Immobilization for Rare Cell Retention During 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 inefficient immobilization during reagent treatment, leading to analysis difficulties and diagnostic challenges.
Innovation Solution
A method and apparatus utilizing a phase change material to immobilize cells on a chamber surface by solidification, preventing cell loss through localized immobilization and stabilization using a bezel unit and thickness adjustment, followed by staining.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional pretreatment methods (cytospin or smear) are used to localize cells on the slide, then cells can be positioned for staining, but 30% or more of cells are lost during reagent treatment and washing
Solution Approach 1:
A droplet-forming element (capillary tube or micropipette) is introduced as an intermediary tool to deliver staining reagents directly to localized cells in a confined droplet space. This intermediary structure prevents reagents from washing away localized cells while enabling effective staining, thereby resolving the contradiction between cell localization and cell retention during reagent treatment
Solution Approach 2:
A coverslip or thin film structure is used to create a confined space over the localized cells. This thin film structure maintains the droplet containing cells and reagents in a stable configuration, preventing cell loss during washing while allowing staining to proceed effectively
2Reliability
If plasma treatment is used to immobilize cells on membrane surface, then cell loss may be reduced, but the method requires expensive equipment and has low reproducibility
Solution Approach 1:
The invention uses simple, inexpensive, disposable items such as capillary tubes or micropipettes as droplet-forming elements instead of expensive plasma treatment equipment. These simple tools can be easily handled and disposed of, providing a cost-effective alternative that maintains high cell retention rates without requiring complex equipment
Solution Approach 2:
The method changes the physical parameters of the staining process by confining cells and reagents in a small droplet volume within a capillary tube or on a coverslip. This parameter change (from bulk solution to confined droplet) enables effective staining with minimal cell loss using simple equipment, avoiding the need for plasma treatment
3Area of stationary object
If cells are widely spread using smear method, then cells can be distributed on the slide, but cell loss occurs during drying and staining processes
Solution Approach 1:
Instead of widely spreading cells across a large slide area, the invention concentrates cells into a localized droplet within a capillary tube or on a small coverslip area. This local quality approach maintains adequate cell distribution within the confined space while preventing cell loss during drying and staining, as the cells remain in a protected droplet environment
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 reduces cell loss to 5% or less, enabling precise analysis and isolation of rare cells, facilitating accurate clinical diagnosis and observation.
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.