Anybody has any experience/pointers as to how best perform whole-cell on a DRG neuron and then lift it from the substrate to continue recording? We are plating the DRG cells in very low poly-L-lysine in glass coverslips. Most of the time we just loose the seal.
How do you lift it? Sudden lift? I used to manipulate first with micro manipulators, like forward-backward-left-right. Repeated several times, then I used macro manipulator to lift it.
Hello, I did this with cortical neurons from brain slice (rat) and for me what worked was to do it *very slowly* it would take me 5-10 min or so to lift the cell with the micromanipulator. You need to add about 20-40 mbar suction to your recording pipette to prevent the membrane from detaching when pulling away. Make sure that your patch pipette is not too large (~6 or 7 MOhm)
I used the nucleated patch technique (aiming to lift the soma, not dendrites/axon of the cells, so the soma would reseal after lifting). I have also done it with primary cortical cells and it works. It should work with DRGs... Good luck!
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Thanks Erkin and Mariana for your answers, will try keeping the suction on!
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