Hi all,

We are exhausting troubleshooting ideas here and I would appreciate new inputs.

We need to record spontaneous firing of the neurons in slices but we are only getting single spikes when injecting currents.

Info:

- I have preliminary data recording spontaneous firing in neurons at the same area. At that time I was using a NMDG solution, holding aCSF, recording aCSF, and pipette solution by Ting et al 2018 (Article Preparation of Acute Brain Slices Using an Optimized N-Methy...

- Here in my lab we tried the same solutions but only had single spikes

- The slices and cells look very healthy and patch easily

- We tried patching different areas of the slice and different slices and only got single spikes.

- Since NMDG can be a problem for firing we tried: 50% NMDG in regular aCSF, 50%NMDG in holding aCSF, only holding aCSF for dissection and slicing - only had single spikes

- We changed the internal solution 3 times - only single spikes

- Today we are trying a sucrose solution and a new aCSF but I am unsure what to expect.

Does anyone have any idea of what could be going on?

Thank you!!!