Single Cell Electroporation
What is Single Cell Electroporation?
In biological research, it is often necessary to introduce foreign macromolecules, such as nucleotides, DNA/RNA, proteins, dyes, and virus particles, into prokaryotic and eukaryotic cells. Electroporation is one of the effective methods to realize macromolecule transportation in or out of cells.
Electroporation, also known as electrotransfection or electropermeabilization, is a process that nanopores form on the cell membrane under the action of the high-intensity electric field, thus the cell membrane permeability increases allowing molecule transportation in or out of cells. It divides into bulk electroporation and single-cell electroporation. Traditionally and commonly used is bulk electroporation, which is commonly used for mass transfer of DNA molecules into a large population of prokaryotic or eukaryotic cells. Single-cell electroporation is a recently developed technique that transports molecules in or out to a single target cell. Compared with bulk electroporation, single-cell electroporation is advantageous in high transfection efficiency, enabling qualitative and quantitative control to target individual cells with little interference with cell viability. Also, this electroporation is used for extraction and analysis of the contents of the single cells, being important to study the distribution and difference of key parameters between different individual cells.
Fig.1 Single cell electroporation. (Santra, 2016)
Applications of Single Cell Electroporation in Neuroscience
Single cell electroporation has emerged as an important tool in genetic study and gene therapy development in neuroscience research. The rapid and precise single neuron electroporation technique realizes analyzing gain and loss of functions of specific genes or proteins. Moreover, single neuron electroporation also has advantages as follows:
- It is available to gene transportation in any species, including those neuronal cell types that cannot be infected by viral vectors.
- It allows to simultaneously transfect multiple genes or plasmids into the neurons.
- It has no or little effect on the electrophysiological properties of the neuron.
- It is easy to be qualitatively and quantitatively controlled.
Based on this single cell electroporation, scientist Haas (Haas, 2001) has successfully developed a micropipette electroporation technique that target transfers a plasmid encoding green fluorescent proteins (GFP) into the brain neuron of intact Xenopus tadpoles. Combining the in vivo time-lapse imaging, the morphology, dendritic arbor branch dynamics, and growth rates could be observed and analyzed in GFP-expressed neurons. Later in 2009, Judkewitz et al. successfully delivered a plasmid DNA into the soma of neocortical neuron of mammalian by single cell electroporation. More recently, single neuron electroporation has been also used for expressing recombinant fluorescent synaptic proteins in target neurons, aiming to in vivo image the synaptic structure and function (Pagès, 2015).
Fig.2 Single-Cell Electroporation Efficiently Transfers Multiple Plasmids into Individual Cells In Vivo. (Haas, 2001)
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References
- Santra, T.S. Tseng, F.G. Electroporation for Single-Cell Analysis. In: Essentials of Single-Cell Analysis. 2016, pp: 55-83.
- Haas, K.; et al. Single-Cell Electroporationfor Gene Transfer In Vivo. Neuron. 2001, 29(3): 583-591.
- Pagès, S.; et al. Single cell electroporation for longitudinal imaging of synaptic structure and function in the adult mouse neocortex in vivo. Frontiers in Neuroanatomy. 2015, 9: 36.
