Cell Monolayer Electroporation Chamber for Lower-Toxicity Pulsing
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Solution Overview
Problem
Existing electroporation methods face challenges with cell heterogeneity, leading to inefficiencies and cellular toxicity due to variations in cell size, membrane composition, and electric field strength requirements, which affect cell viability and cost-effectiveness.
Innovation Solution
The use of a sample container with an insulator chamber and electrodes to form a compact cell monolayer, combined with a pulse generator to deliver controlled electrical pulses, minimizes cell-to-cell interactions and reduces electric field strength, enhancing electroporation efficiency and cell survival.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If high field strength is used for electroporation, then electroporation efficiency is improved, but cellular toxicity increases
Solution Approach 1:
The patent applies local quality by creating a non-uniform electric field distribution where the field strength is concentrated at specific locations (between closely spaced electrodes) while remaining lower elsewhere. This allows high field strength to be applied locally to achieve efficient electroporation without subjecting the entire cell population to uniformly high field strengths that cause toxicity.
Solution Approach 2:
The patent segments the electroporation process by using multiple closely spaced electrodes instead of a single pair, creating multiple localized high-field regions. This segmentation allows different portions of the cell suspension to experience different field strengths, improving overall efficiency while reducing average toxicity.
2Productivity
If high voltage is applied to electroporate smaller cells, then electroporation efficiency for smaller cells is improved, but larger cells are irreversibly damaged
Solution Approach 1:
The patent creates local quality variations in the electric field by using electrodes at different positions and orientations. Smaller cells located in specific regions experience higher field strengths suitable for their electroporation requirements, while larger cells in other regions experience lower field strengths that prevent irreversible damage.
Solution Approach 2:
The patent employs dynamic control of the electric field through programmable pulse generators that can adjust voltage and pulse duration based on real-time conditions. This dynamic adjustment allows the system to optimize field strength for different cell sizes present in the suspension, improving efficiency for smaller cells while protecting larger cells.
3Ease of manufacture
If uniform electric field is applied to heterogeneous cell population, then simplicity of process is maintained, but electroporation efficiency varies across cells
Solution Approach 1:
The patent segments the electric field application into multiple localized zones using closely spaced electrodes. This segmentation creates a more complex field distribution pattern that naturally adapts to heterogeneous cell populations, improving efficiency consistency without requiring complex external control systems.
Solution Approach 2:
The patent enables the electric field to self-adjust to the cell population heterogeneity through the geometric arrangement of multiple electrodes. The field distribution automatically adapts to the spatial distribution of cells, improving electroporation efficiency consistency without requiring active sensing or control mechanisms.
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 increases electroporation efficiency by up to 67.3% for uniform cell sizes and reduces cellular toxicity, lowering power requirements and manufacturing costs, while maintaining consistent molecular delivery across cells.
Implementation Method 1
Electroporation is a widely-used method for permeabilization of cell membranes by temporary generation of membrane pores with electrical stimulation
Implementation Method 2
Electroporation is a widely-used method for permeabilization of cell membranes by temporary generation of membrane pores with electrical stimulation
Implementation Method 3
The insulator chamber is configured to contain at least one cell monolayer
Implementation Method 4
the heat generated by the electric current may harm the cells
Implementation Method 5
The cell suspension is then placed in a rectangular cuvette embedded with two flat electrodes for an electrical discharge
Data Source
AI summary
An apparatus for electroporation of biological cells is provided. The apparatus includes a sample container having an insulator chamber for holding the cells. The sample container has a first electrode and a second electrode to provide electrical connection for electroporation. The insulator chamber is configured to contain at least one cell monolayer. The apparatus also includes a pulse generator that can generate a predetermined pulse for electroporation of the cells.