Here is a preliminary working template based on protocols of the first batch of the AIBS cell type database (see http://help.brain-map.org/download/attachments/8323525/CellTypes_Ephys_Overview.pdf?version=2&modificationDate=1508180425883&api=v2 in the section Appendix: Stimulus Details). Deviation are marked with ! . Please follow the template protocol as closly as possible/sensible given the differences in aquisition equipment:
Different "stimulus segments"/epoches are separated by "\".
0. General features
__a. The prestimulus intervall of all recorded sweeps should have the same structure including a small and short hyperpolarizing! test pulse:
40 ms 0 pA Step (no increment over sweeps) \ 10 ms square pulse step to -25 pA (no increment over sweeps) \ 450 ms 0 pA Step (no increment over sweeps)
__b. Sampling frequency for continous amplifiers is 50kHz. In discontinous amplifiers as high as sensible/possible (at least 25kHz)!
__c. except when measuring the membrane potential within the first minute(s) after breakthrough, a holding current is applied to keep the baseline from drifting. The readjustment can be done either manually or automatically using algorithms as provided in MIES.
1. Ramp (optional)
__a. 3 repetetions of single sweep of \ 20 s 0-to-500 pA ramp (slope: 25 pA/s) terminated manually after a series of action potentials are acquired) with an interstimulus\interrepettion intervall of >10 seconds.
2. Long Square Pulse
__a. 1 repetetion of sweeps \ 1000 ms Step to -110 pA (+20 pA increment over sweeps until first supratheshold level + 160/260 pA [depending on input resistance] are reached) \ 6500 ms 0 pA Step (no increment over sweeps)
Start-to-Start Interval: 10 sec for all! AIBS actually uses 10 sec for subthreshold 17 sec for suprathreshold, but imho too complex for now
Side note: in rare cases with very low input resistance AIBS starts with current injections at -190 or -210 with a higher increment
__b. 2-3 repetetion of a single sweep of \ 1000 ms "rheobase" level pA Step \ 6500 ms 0 pA Step (no increment over sweeps)
Comment:The AIBS makes sure that they have 3 repetetions of the "rheobase" level spike for each cell. How many extra sweeps depends on if 2a contains a sweep with a single spike or something close to it (not possible if intrinsic bursting etc.), which technically is not the exact definitation of rheobase so it is in "" here.
3. Short Square Pulse
__a. 1 repetetion of sweeps of \ 3 ms +100 pA (+10 pA increment over sweeps ) \ 1497 ms 0 pA (no increment over sweeps) \ Start-to-Start Interval: 4 sec. AIBS actually uses 4 sec for subthreshold 5 sec for suprathreshold, but imho too complex for now
__b. at least 3 more repetitions of a single sweep of \ 3 ms first supratreshold level (no increment over sweeps) \ 1497 ms 0 pA (no increment over sweeps) \ Start-to-Start Interval: 5 sec
__c. if 3b did not produce an action potential at every repetition repeat 3b with 10 pA higher current step