Phototagging Agent for Selective Cell Isolation
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for isolating cells from various samples, such as two-dimensional cell cultures and three-dimensional tissues, lack versatility and efficiency while often causing cell damage, and there is a need for improved techniques to selectively isolate target cells without harming them.
Innovation Solution
A method using a phototagging agent that changes fluorescence response upon activation with photo-activating light, allowing for selective isolation of target cells based on fluorescence differences, combined with a microscope system capable of imaging and directing activating light to specific cells, and a chemical compound comprising a caged fluorophore conjugated with a photosensitizer for enhanced uncaging efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional cell isolation methods are used, then cells can be separated from samples, but cell damage occurs and versatility across different sample types is limited
Solution Approach 1:
The patent employs fluorescence-based optical tagging where target cells are selectively labeled with fluorescent markers. This allows non-contact identification and isolation of specific cells based on their fluorescence signal, avoiding mechanical stress that causes cell damage while maintaining versatility across 2D cultures, 3D tissues, and biopsy samples
Solution Approach 2:
The invention replaces conventional mechanical isolation methods (such as micromanipulation or physical sorting) with an optical-based system. The microscope system with beam patterning optics directs photo-activating light to specific target cells, and fluorescently labeled cells are isolated based on their optical signal rather than physical manipulation, thereby preventing cell damage while handling diverse sample types
2Productivity
If photochemical crosslinkers are used for cell selection, then cells can be crosslinked to extracellular matrix, but the process lacks efficiency and versatility for different sample types
Solution Approach 1:
The patent describes a universal cell isolation system that can handle multiple sample types including two-dimensional cell cultures, three-dimensional tissues, biopsies, and spheroid cultures. The microscope system with beam patterning optics and phototagging approach provides a unified methodology applicable across all these diverse sample formats, eliminating the need for sample-type-specific protocols and improving overall productivity
Solution Approach 2:
The invention utilizes photo-activatable fluorophores that undergo parameter changes upon light activation. The fluorophores transition from non-fluorescent to fluorescent states when exposed to specific wavelengths, enabling selective marking of target cells. This parameter change mechanism provides efficient and versatile cell selection across different sample types without requiring conventional crosslinking chemistry
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 enables efficient and selective isolation of target cells with minimal damage, allowing for precise identification and separation of cells in different sample types, with improved uncaging efficiency and longer-lasting fluorescence changes for easier identification.
Implementation Method 1
The identified target cell in the sample is selectively irradiated with photo-activating light for selectively activating the phototagging agent in the target cell to change its fluorescence response
Implementation Method 2
The irradiated target cell is isolated from other cells in the sample based on a difference in its fluorescence response compared to non-activated phototagging agent in the other cells
Data Source
AI summary
According to one method, a sample with cells contains a phototagging agent. The sample is imaged to identify at least one target cell to be isolated. The identified target cell in the sample is selectively irradiated with photo-activating light for selectively activating the phototagging agent in the target cell to change its fluorescence response. The irradiated target cell is isolated from other cells in the sample based on a difference in its fluorescence response compared to non-activated phototagging agent in the other cells. Further aspects are directed to a corresponding microscope system and chemical compound for use as the phototagging agent.