Microfluidic Live Cell Sampling via Electroporation and Nanochannels
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Conventional methods for investigating single biological cells rely on destructive lysis to extract and analyze internal contents, providing information only at end points and failing to capture dynamic cellular processes.
Innovation Solution
A microfluidic technology (Live Cell Analysis Device or LCAD) uses electric pulses to temporarily open pores in the cell membrane, allowing non-destructive extraction and analysis of intracellular contents into nano-liter chambers, enabling dynamic cellular process investigation and perturbation with external molecules.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional lysis methods are used to extract intracellular contents, then analysis can be performed, but the cell membrane is destroyed and only endpoint information is obtained
Solution Approach 1:
The patent applies periodic electroporation pulses to temporarily open cell membrane pores in a controlled, repeating manner. Each pulse sequence opens pores for a brief period to allow intracellular content extraction, then closes them to restore membrane integrity. This periodic action enables multiple sampling timepoints from the same live cells without permanent damage, resolving the contradiction between analysis capability and cell viability.
2Loss of time
If electric pulses are applied to open membrane pores for content extraction, then intracellular contents can be analyzed at multiple timepoints, but the cell membrane is temporarily disrupted
Solution Approach 1:
The patent carefully controls electroporation parameters including pulse voltage (e.g., 10-30V), pulse duration (e.g., 10-100 microseconds), pulse frequency, and buffer composition to optimize the temporary membrane pore opening. By adjusting these parameters, the system achieves sufficient content extraction for temporal analysis while minimizing membrane disruption and ensuring rapid membrane recovery, thus resolving the contradiction between temporal resolution and membrane integrity.
3Productivity
If cells are cultured in conventional formats, then cell growth is supported, but dynamic cellular processes cannot be monitored over time without cell destruction
Solution Approach 1:
The patent merges conventional cell culture capabilities with electroporation-based content extraction in an integrated microfluidic platform. The system combines cell culture chambers with electroporation electrodes and content collection reservoirs, allowing simultaneous cell growth maintenance and periodic intracellular content sampling. This integration enables long-term culture of live cells while capturing dynamic cellular processes at multiple timepoints, resolving the contradiction between culture capacity and dynamic information acquisition.
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
Enables non-destructive, multipoint analysis of intracellular contents, preserving cell functionality, and allows for long-term culture and monitoring of cells, facilitating studies of dynamic cellular processes and responses to external inputs.
Implementation Method 1
the membrane is configured to allow diffusion of substances across the membrane into the layer of extraction chambers
Implementation Method 2
uses electric pulses to open pores temporarily in the cell membrane, allowing for their internal contents to leak out
Data Source
AI summary
A cell analysis system includes a multi-layer microfluidic device that includes a layer of microfluidic channels, a layer of microwells, a membrane with nanochannels, and a layer of extraction chambers. The microwells and the membrane are configured to allow culturing of cells that are adhered to the membrane or suspended in the microwells, and the membrane is configured to allow diffusion of substances across the membrane into the layer of extraction chambers. The cell analysis system includes a top conductive layer and a bottom conductive layer on the opposite sides of the multi-layer microfluidic device. The cell analysis system also includes a function generator configured to apply an electroporation pulse between the top conductive layer and the bottom conductive layer.