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Internalization and Observation of Fluorescent Biomolecules in Living Microorganisms via Electroporation
Published on: February 8, 2015
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A live mammalian cells electroporation array for on-chip immunofluorescence
Marta Maschietto1, Stefano Girardi1, Onelia Gagliano2
1Department of Biomedical Sciences, Section of Physiology, University of Padua, via F. Marzolo 3, 35131 Padua, Italy.
Journal of Immunological Methods
|December 25, 2023
Summary
This study shows in situ electroporation efficiently delivers fluorescent markers into cells using microelectrodes. This technique enables detection of intracellular proteins without chemical fixation, applicable to various cell types and targets.
Area of Science:
- Biotechnology
- Cell Biology
- Neuroscience
Background:
- Intracellular protein detection typically uses immunofluorescence on fixed cells.
- Chemical fixation can alter cellular structures and protein epitopes.
- Electroporation offers a non-chemical method for introducing molecules into cells.
Purpose of the Study:
- To demonstrate efficient in situ electroporation for intracellular protein detection.
- To utilize thin film silicon dioxide (SiO2) capacitive microelectrodes for electroporation.
- To validate the technique in mammalian cell lines and primary neurons.
Main Methods:
- In situ electroporation on SiO2 capacitive microelectrodes.
- Delivery of fluorescent markers and antibodies into live cells.
- Detection of intracellular proteins, exemplified by actin.
Main Results:
- High efficiency of in situ electroporation achieved.
- Successful delivery of markers and antibodies into mammalian cells and primary neurons.
- Demonstrated detection of intracellular proteins like actin.
Conclusions:
- In situ electroporation on microelectrodes is an effective method for intracellular protein detection.
- This technique avoids chemical fixation, preserving cellular integrity.
- The method is versatile and can be extended for detecting various target proteins.
