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A Microfluidic Chip for the Versatile Chemical Analysis of Single Cells
Published on: October 15, 2013
Single-cell electroendocytosis on a micro chip using in situ fluorescence microscopy
Ran Lin1, Donald C Chang, Yi-Kuen Lee
1Bioengineering Graduate Program, The Hong Kong University of Science and Technology, Clear Water Bay, Hong Kong.
Biomedical Microdevices
|July 30, 2011
Summary
Electroendocytosis (EED) uses electric fields for cellular delivery, forming vesicles instead of pores. This study explores EED in HeLa cells, revealing biphasic efficiency and distinct mechanisms from electroporation.
Area of Science:
- Biotechnology
- Cell Biology
- Bioengineering
Background:
- Electroendocytosis (EED) is an electric field-induced cellular uptake method.
- Unlike electroporation (EP), EED utilizes endocytotic vesicles for molecule delivery.
- Understanding EED mechanisms is crucial for advanced drug and biomolecule delivery systems.
Purpose of the Study:
- To comprehensively investigate electroendocytosis (EED) in HeLa cells using a microchip.
- To visualize and analyze the endocytotic vesicles formed during EED.
- To characterize EED efficiency based on electric field parameters and elucidate its mechanism.
Main Methods:
- Utilized a microchip for electroendocytosis experiments on HeLa cells.
- Employed FM4-64 fluorescent dye and in situ fluorescence microscopy for vesicle visualization.
- Quantified intracellular fluorescent intensity in over 2,000 single cells to assess molecule uptake.
- Investigated EED mechanisms using cytoskeleton inhibitors.
Main Results:
- Visualized and investigated EED-induced endocytotic vesicles in HeLa cells.
- Determined EED efficiency as a function of electric field strength, pulse duration, and total treatment time.
- Observed biphasic characteristics in EED efficiency concerning electric field strength.
- Demonstrated EED mechanisms distinct from electroporation through cytoskeleton inhibitor experiments.
Conclusions:
- EED is a viable method for cellular delivery, distinct from EP.
- EED efficiency is dependent on electric field parameters, exhibiting biphasic behavior.
- The study provides foundational insights into the time-dependent mechanism of EED at the single-cell level, paving the way for on-chip investigations.
