Optical Tweezer Cell Lifting in PEG-Grafted OSTE+ Microwells
Solution Overview
Problem
Current methods for single B cell selection and sorting are inefficient, leading to viability loss and contamination, and require faster screening tools with higher efficiency rates for high-throughput analysis.
Innovation Solution
Integration of an optical tweezer system with a PEG-grafted OSTE+ microwell array, utilizing a microfluidic device with branched channels and a unique droplet-based cell retrieval system to minimize interaction times and prevent cell loss, allowing for the selective trapping and lifting of viable B cells without affecting their viability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional FACS sorting and single cell sequencing methods are used, then single B cell selection can be achieved, but the process is inefficient and results in viability loss
Solution Approach 1:
The invention divides the cell population into individual single-cell compartments within a microarray, allowing parallel processing of multiple cells simultaneously. This segmentation enables high-throughput screening while maintaining individual cell integrity and viability, resolving the contradiction between efficiency and viability loss
Solution Approach 2:
The patent replaces conventional mechanical FACS sorting with an optical tweezer-based system that uses optical forces to manipulate and select cells. This substitution eliminates the need for physical shearing and mechanical stress that cause viability loss, achieving both high efficiency and cell survival
2Productivity
If faster screening tools are implemented, then throughput increases, but cell loss and contamination occur
Solution Approach 1:
The invention embeds multiple functional components within a single microarray platform: cell trapping wells, optical tweezer interaction zones, and collection channels are integrated into one structured system. This nesting enables high-throughput processing while containing cells throughout the entire workflow, preventing cell loss and contamination
Solution Approach 2:
The patent introduces a microfluidic channel system as an intermediary between cell introduction and collection, providing controlled fluid flow that guides selected cells through the system. This intermediary structure enables fast screening throughput while maintaining precise cell control and preventing accidental cell loss
3Measurement precision
If optical tweezers are used to lift cells from wells, then single cell selection is achieved, but interaction time increases
Solution Approach 1:
The invention pre-positions cells in microwell locations that are optimally accessible to optical tweezers, and pre-configures the microarray structure to enable rapid cell ejection. This preliminary arrangement allows precise single-cell selection with minimized interaction time, resolving the contradiction between selection precision and time efficiency
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 system enables high-throughput screening and efficient collection of viable B cells with a positive signal, maintaining cell viability and reducing contamination, allowing for further analysis such as single cell sequencing.
Implementation Method 1
single cell lifting of the yet viable cell from the well by an optical tweezer
Implementation Method 2
PEG-grafted OSTE+ microwell array, this array being grafted with methacrylated polyethylene glycol
Data Source
AI summary
In general the present invention concerns 1) single cell trapping of a viable cell in separate well from a plurality of wells in an array of wells, 2) single cell analysis for the selected cell and 3) single cell lifting of the yet viable cell from the well by an optical tweezer. Furthermore resent invention concerns a cell trap and lift device for B lymphocytes, the device comprising an array of wells in in polymer matrix comprising an off-stoichiometry thiol-ene polymer of the group consisting of off-stoichiometry thiol-ene (OSTE) and off-stoichiometry thiol-ene-epoxy (OSTE+) or a combination thereof that have been grafted with methacrylated polyethylene glycol (methoxy polyethylene glycol methacrylate or (M-PEG-M)) of a number average molecular weight of Mn 2000. It furthermore concerns using the B lymphocyte trap and lift device for trapping single B lymphocyte cells in wells of the device of present invention and lifting said cell from the cell trapping well by optical tweezers, preferably single beam tweezers.