Single-Cell Electroporation of Neurons
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Single-Cell Electroporation of Neurons. / Wiegert, J Simon; Gee, Christine E; Oertner, Thomas G.
in: Cold Spring Harbor protocols, Jahrgang 2017, Nr. 2, 01.02.2017, S. pdb.prot094904.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - Single-Cell Electroporation of Neurons
AU - Wiegert, J Simon
AU - Gee, Christine E
AU - Oertner, Thomas G
N1 - © 2017 Cold Spring Harbor Laboratory Press.
PY - 2017/2/1
Y1 - 2017/2/1
N2 - Single-cell electroporation allows the transfection of a small number of neurons in an organotypic culture with a single plasmid or a defined mixture of plasmids. Desired protein expression levels can vary depending on the experimental goals (e.g., high expression levels are needed for optogenetic experiments); however, when too much protein is expressed, cellular toxicity and cell death may arise. To a large degree, protein expression can be controlled by adjusting the concentration of plasmid DNA in the electroporation pipette. Here, we present a protocol for transfecting individual neurons in hippocampal slice cultures by electroporation. Essentially, a patch-clamp setup is required that includes an upright microscope with infrared differential interference contrast or Dodt contrast with a camera and a specialized amplifier that is able to deliver large-voltage pulses to the electroporation pipette.
AB - Single-cell electroporation allows the transfection of a small number of neurons in an organotypic culture with a single plasmid or a defined mixture of plasmids. Desired protein expression levels can vary depending on the experimental goals (e.g., high expression levels are needed for optogenetic experiments); however, when too much protein is expressed, cellular toxicity and cell death may arise. To a large degree, protein expression can be controlled by adjusting the concentration of plasmid DNA in the electroporation pipette. Here, we present a protocol for transfecting individual neurons in hippocampal slice cultures by electroporation. Essentially, a patch-clamp setup is required that includes an upright microscope with infrared differential interference contrast or Dodt contrast with a camera and a specialized amplifier that is able to deliver large-voltage pulses to the electroporation pipette.
U2 - 10.1101/pdb.prot094904
DO - 10.1101/pdb.prot094904
M3 - SCORING: Journal article
C2 - 28148856
VL - 2017
SP - pdb.prot094904
JO - Cold Spring Harbor protocols
JF - Cold Spring Harbor protocols
SN - 1559-6095
IS - 2
ER -