We are interested in doing in vivo electroporation in neonatal mouse brain. For this we are looking for a suitable electrode. The tweezertrodes available commercially are ideal for this but they seem to be way overpriced. I am wondering if there is anyone who build their own electrodes and how you do it. Any suggestions will be welcome! Thank you!
Dear Xiwei,
I have been using single cell electroporation in brain slices and whole-mount retinas. For this purpose I have been using just patch-electrodes with 3-4 megaohm resistances (see my 2008 and 2013 electroporation methodological papers). I have been filling cells with dyes, and more recently with mRNA using single cell electroporation. You can build your own glass electrodes depending on where and how deep you need to deep into the brain. You can basically pull electrodes with long shunks and then break the tips to the size that you prefer, probably ~ 1 megaohm or 2-3 micron, then fire polish the tip. You go the region of your interest, depending on how many and how much you want to electroporate cells, apply voltage or current pulses (depending on the size of your molecule you are trying to electroporate). For single cells I typically use ~0.5 nA or 50-100 mV for 0.5 s at 1 Hz (total of 4-5 mins). If your molecules are large than you will need to go about several folds higher, so that you break the membrane briefly but large enough to pass your molecule into the cytoplasm. This is the basics that you can start with and then you will need to experiment to find ideal parameters for your own experiment.
best wishes,
Refik
- Badges
- Science topic
- Similar topics
- Engineering
- Electrical Engineering
- Electrodes