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Updated: Dec 27, 2025
Single-Cell Electroporation across Different Organotypic Slice Culture of Mouse Hippocampal Excitatory and Class-Specific Inhibitory Neurons
Published on: October 6, 2020
A highly efficient method for single-cell electroporation in mouse organotypic hippocampal slice culture
David G Keener1, Amy Cheung2, Kensuke Futai3
1Brudnick Neuropsychiatric Research Institute, Department of Neurobiology, University of Massachusetts Medical School, 364 Plantation Street, LRB-706, Worcester, MA, 01605-2324, USA; Graduate Program in Neuroscience, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
Background:
Exogenous gene introduction by transfection is one of the most important approaches for understanding the function of specific genes at the cellular level. Electroporation has a long-standing history as a versatile gene delivery technique in vitro and in vivo. However, it has been underutilized in vitro because of technical difficulty and insufficient transfection efficiency.
New Method:
We have developed an electroporation technique that combines the use of large glass electrodes, tetrodotoxin-containing artificial cerebrospinal fluid and mild electrical pulses. Here, we describe the technique and compare it with existing methods.
Results:
Our method achieves a high transfection efficiency (∼80 %) in both excitatory and inhibitory neurons with no detectable side effects on their function. We demonstrate this method is capable of transferring at least three different genes into a single neuron. In addition, we demonstrate the ability to transfect different genes into neighboring cells.
Comparison With Existing Methods:
The majority of existing methods use fine-tipped glass electrodes (i.e. > 10 MΩ) and apply high voltage (10 V) pulses with high frequency (100 Hz) for 1 s. These parameters contribute to practical difficulties thus lowering the transfection efficiency. Our unique method minimizes electrode clogging and therefore procedure duration, increasing transfection efficiency and cellular viability.
Conclusions:
Our modifications, relative to current methods, optimize electroporation efficiency and cell survival. Our approach offers distinct research strategies not only in elucidating cell-autonomous functions of genes but also for assessing genes contributing to intercellular functions, such as trans-synaptic interactions.
