Cell Sorting Using Hydrogel Curing and Fluorescent Markers
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Solution Overview
Problem
Current rare cell detection and sorting systems face challenges in achieving high purity, high throughput, high cell recovery rate, and low cell damage due to the rarity and fragility of cells such as circulating tumor cells and fetal nucleated red blood cells.
Innovation Solution
A cell sorting method using an array chip and a hydrogel solution, where fluorescent markers are used to bind with target cells, and the mixture is distributed and cured on the chip to fix cells, allowing for precise fluorescence image analysis and isolation of target cells with minimal damage.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional cell sorting methods are used, then cell detection and sorting can be performed, but cell damage occurs and cell recovery rate is low
Solution Approach 1:
The patent changes the physical state of the hydrogel from liquid to solid through temperature change (cooling below gelation temperature) or photoirradiation (for photosensitive hydrogel), thereby fixing the cells in position without mechanical stress or enzymatic treatment that could damage them. This parameter change enables gentle cell fixation and high recovery rate.
Solution Approach 2:
The hydrogel undergoes phase transition from liquid to solid state, which allows the cells to be fixed in their positions without causing damage. The phase transition provides a gentle yet effective method to immobilize cells for subsequent analysis while maintaining cell viability and enabling high recovery rates.
2Measurement precision
If traditional sorting methods are used, then cells can be separated, but sorting purity and accuracy are insufficient
Solution Approach 1:
The patent uses fluorescent markers that emit different colors or fluorescence intensities when bound to target cells. This optical property enables precise identification and differentiation of target cells from non-target cells through fluorescence imaging, thereby achieving high sorting accuracy and purity.
Solution Approach 2:
The patent creates a spatial copy or map of cell positions and fluorescence characteristics through imaging before isolation. This allows for precise identification and selection of target cells based on their fluorescence signal, ensuring high sorting accuracy and purity without physical contact that could cause damage.
3Stability of the object's composition
If cells are fixed using conventional methods, then position can be stabilized, but cell damage occurs
Solution Approach 1:
The hydrogel acts as an intermediary medium between the liquid cell suspension and the solid support surface. It provides a gentle transition state where cells can be embedded and fixed without direct contact with harsh fixing agents or mechanical forces, thereby stabilizing cell positions without causing damage.
Solution Approach 2:
The patent changes the physical state of the hydrogel from liquid to solid through temperature change or photoirreversal, thereby fixing the cells in position without mechanical stress or enzymatic treatment that could damage them. This parameter change enables gentle cell fixation and high recovery rate.
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 method improves the accuracy and purity of cell sorting while reducing cell damage and achieving a high cell recovery rate, as demonstrated by the use of temperature-sensitive or photosensitive hydrogel solutions that fix cells without causing significant harm.
Implementation Method 1
Fluorescent markers are added to the cell mixture to allow the fluorescent markers to bind with the target cells
Implementation Method 2
The step of curing the mixture solution includes: a cooling process is performed to cure the mixture solution, positions of the target cells and the non-target cells are fixed
Implementation Method 3
The step of curing the mixture solution includes: the first region is irradiated with ultraviolet light to cure the mixture solution located in the first region, so as to fix the target cells
Data Source
AI summary
Cell sorting methods are disclosed. A blood sample is provided. The blood sample is centrifuged to obtain a cell mixture having target cells and non-target cells. Fluorescent markers are added to the cell mixture to allow the fluorescent markers to bind with the target cells. A hydrogel solution is added and mixed with the cell mixture to form a mixture solution. The mixture solution is distributed to cell wells of an array chip, and the target cells and non-target cells in the cell mixture are spread on the bottom of the cell wells in a substantially monolayer manner. The mixture solution is cured. A fluorescence image analysis is performed to select the target cells that are bound with fluorescent markers and emit fluorescence, and the selected target cells are isolated.