Microfabricated Device for Low-Voltage Single Cell Electroporation
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Solution Overview
Problem
Current methods for single cell electroporation lack control and efficiency, as they require high voltages and are not suitable for reversible permeabilization, making it difficult to manipulate individual cells effectively.
Innovation Solution
A microfabricated device with lateral cell trapping junctions and microfluidic channels allows for focused electric fields and low-voltage electroporation, enabling reversible electroporation and electrofusion of cells with precise control over transmembrane potential and compound delivery.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If bulk electroporation is used to deliver DNA to mammalian cells, then transfection efficiency is improved, but control over individual cells is lost and suboptimal parameters result
Solution Approach 1:
The invention divides the bulk cell population into individual single cells, each treated separately in a microfabricated device. This segmentation enables precise control over each cell's electroporation parameters while maintaining high throughput, resolving the contradiction between bulk transfection efficiency and individual cell control.
Solution Approach 2:
The microfabricated device with micropores acts as an intermediary structure that holds individual cells in place and delivers electroporation pulses with precise spatial and temporal control. This intermediary enables both high efficiency and individual cell control simultaneously.
2Reliability
If high voltages are applied for electroporation, then membrane permeabilization is achieved, but reversible electroporation becomes difficult and cell damage increases
Solution Approach 1:
The invention applies electroporation voltage locally across the cell membrane through the micropore structure, concentrating the electric field precisely where needed. This localized application achieves reliable membrane permeabilization while minimizing overall voltage exposure and reducing cell damage, enabling reversible electroporation.
Solution Approach 2:
The microfabricated device enables precise control of electroporation parameters including voltage, pulse duration, and waveform. By optimizing these parameters within a controlled range rather than using high voltages, the system achieves effective permeabilization while avoiding cell damage and enabling reversibility.
3Ease of operation
If single cell electroporation is performed with carbon fiber microelectrodes, then individual cell control is achieved, but very high voltages are required and the system becomes complex
Solution Approach 1:
The invention replaces the mechanical carbon fiber microelectrode positioning system with a microfabricated device where cells are passively held in micropores. This substitution eliminates the need for high voltages and complex positioning mechanisms, achieving individual cell control with lower energy requirements.
4Productivity
If microfabricated chips are used for single cell electroporation, then control and efficiency are improved, but device complexity increases
Solution Approach 1:
The microfabricated device integrates multiple functions including cell trapping, electroporation, and compound delivery into a single unified structure. This multi-functionality achieves high electroporation efficiency without proportionally increasing device complexity, as one structure serves multiple purposes.
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 device achieves efficient and reversible electroporation of single cells using low voltages, allowing for controlled introduction of compounds and monitoring of cellular responses, while also enabling cell fusion and high-throughput screening.
Implementation Method 1
This can be accomplished via electroporation, a technique using high electric fields to induce structural rearrangements of the cell membrane. Pores are formed when the transmembrane potential exceeds the dielectric break-down voltage of the membrane (about 0.2-1.5V).
Implementation Method 2
Pores are formed when the transmembrane potential exceeds the dielectric break-down voltage of the membrane
Data Source
AI summary
Method and systems provide improved cell handling in microfluidic systems and devices using lateral cell trapping and methods of fabrication of the same that allow for selective low voltage electroporation and electrofusion.