IUE was performed as previously described (Bonnefont et al., 2019 (link)). Briefly, timed-pregnant mice were anesthesized with a ketamine/xylazine mixture at E13.5, and each uterus was exposed under sterile conditions. Plasmid solutions containing 1-2 mg/ml of DNA were injected into the lateral ventricles of the embryos using a heat-pulled capillary. Electroporation was performed using tweezers electrodes (Nepa Gene) connected to a BTX830 electroporator (five pulses of 25 V for 100 ms with an interval of 1s). Embryos were placed back into the abdominal cavity, and mice were sutured and placed on a heating plate until recovery. Embryos were collected 24 or 48 hours after IUE and perfused transcardiacally with ice-cold 4% paraformaldehyde. Brains were dissected and soaked in 4% paraformaldehyde overnight at 4 °C. They were then washed in PBS for 24 h then soaked in 30% sucrose solution overnight for cryopreservation. Brains were included in Tissue-TEK OCT compound (Sakura) at -20 °C overnight then stored at -80 °C. After equilibration the blocs were sectioned with 20-μm thickness with a cryostat (Leica).
Tweezers electrodes
Manufactured by Nepa Gene
Tweezers electrodes are a type of laboratory equipment used for handling and manipulating small or delicate samples. They feature two narrow, pointed tips that can be used to grasp, move, or apply electrical connections to various materials or components. The core function of tweezers electrodes is to provide a precise and controlled method for interacting with small-scale samples or devices.
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Lab products found in correlation
2 protocols using tweezers electrodes
1
In Utero Electroporation of Mouse Embryos
2
In Utero Electroporation of Mouse Embryos
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In utero electroporation was performed as previously described (Bonnefont et al 2019).
Briefly, timed-pregnant mice were anesthesized with a ketamine/xylazine mixture at E13.5, and each uterus was exposed under sterile conditions. Plasmid solutions containing 1-2 mg/ml of DNA were injected into the lateral ventricles of the embryos using a heat-pulled capillary. Electroporation was performed using tweezers electrodes (Nepa Gene) connected to a BTX830 electroporator (five pulses of 25 V for 100 ms with an interval of 1s). Embryos were placed back into the abdominal cavity, and mice were sutured and placed on a heating plate until recovery. Embryos were collected 24 or 48 hours after in utero electroporation and perfused transcardiacally with ice-cold 4% paraformaldehyde. Brains were dissected and soaked in 4% paraformaldehyde overnight at 4 °C. They were then washed in PBS for 24 h then soaked in 30% sucrose solution overnight for cryopreservation. Brains were included in Tissue-TEK OCT compound (Sakura) at -20 °C overnight then stored at -80 °C. After equilibration the blocs were sectioned with 20-µm thickness with a cryostat (Leica).
Briefly, timed-pregnant mice were anesthesized with a ketamine/xylazine mixture at E13.5, and each uterus was exposed under sterile conditions. Plasmid solutions containing 1-2 mg/ml of DNA were injected into the lateral ventricles of the embryos using a heat-pulled capillary. Electroporation was performed using tweezers electrodes (Nepa Gene) connected to a BTX830 electroporator (five pulses of 25 V for 100 ms with an interval of 1s). Embryos were placed back into the abdominal cavity, and mice were sutured and placed on a heating plate until recovery. Embryos were collected 24 or 48 hours after in utero electroporation and perfused transcardiacally with ice-cold 4% paraformaldehyde. Brains were dissected and soaked in 4% paraformaldehyde overnight at 4 °C. They were then washed in PBS for 24 h then soaked in 30% sucrose solution overnight for cryopreservation. Brains were included in Tissue-TEK OCT compound (Sakura) at -20 °C overnight then stored at -80 °C. After equilibration the blocs were sectioned with 20-µm thickness with a cryostat (Leica).
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