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The Fabrication and Operation of a Continuous Flow, Micro-Electroporation System with Permeabilization Detection
Published on: January 7, 2022
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Continuous microfluidic flow-through protocol for selective and image-activated electroporation of single cells.
Felix Pfisterer1, Neus Godino1, Tobias Gerling1
1Fraunhofer Institute for Cell Therapy and Immunology IZI, Branch Bioanalytics and Bioprocesses IZI-BB Am Muehlenberg 13 14476 Potsdam Germany michael.kirschbaum@izi-bb.fraunhofer.de.
RSC Advances
|June 29, 2023
Summary
This study introduces a microfluidic method for targeted cell electroporation. It selectively modifies specific cells in mixed populations using real-time image analysis, improving molecular delivery and cell analysis.
Area of Science:
- Biotechnology
- Cell Biology
- Microfluidics
Background:
- Electroporation is vital for molecular delivery and cell manipulation.
- Current bulk electroporation methods lack selectivity for specific cell subpopulations.
- Existing selective electroporation requires pre-sorting or complex single-cell technologies.
Purpose of the Study:
- To develop a microfluidic system for real-time, selective electroporation of target cells within heterogeneous samples.
- To enable precise molecular delivery or extraction from identified single cells.
- To overcome limitations of bulk electroporation in complex biological mixtures.
Main Methods:
- A microfluidic chip integrating dielectrophoretic cell focusing and real-time microscopic image analysis.
- Image processing algorithms for classifying cells based on fluorescence and transmitted light.
- A poration electrode system that delivers electrical pulses exclusively to identified target cells.
Main Results:
- Selective electroporation of green-fluorescent target cells in a mixed green/blue fluorescent sample.
- >90% specificity in targeting desired cells.
- Average poration rates exceeding 50% with a throughput of up to 7200 cells per hour.
Conclusions:
- The developed microfluidic protocol enables highly specific electroporation of target cells in real-time.
- This technology offers a significant advancement for selective cell manipulation in heterogeneous samples.
- It provides a foundation for advanced applications in cell biology and biotechnology requiring precise single-cell processing.
