Hi everybody,
I have some problems. Today I decided to clean my setup disconnecting and reconnecting some cables to ameliorates the noise. I disconnected the digidata from the PC removing the USB cables but after the clean operation I connected all as was. Now the problem is that, when I put my electrode in the bath in the membrane test I don't have the classic graph to study the properties of the electrode. I don't know why.
If you can help me to solve this problem I'll be grateful.
I have a digidata 1440 mod. and a multiclamp 700B as amp and pClamp 10 as software.
Thanks
Hi Luca,
There are many possibilities for your problem. I will discard the wrong connection of the cables, assuming that you already checked that. The first possibility is that the recording loop is broken, ie either your recording electrode is not correctly connected or your ground electrode is out of the bath or not connected. Another possibility is that when you restarted your computer your digidata was off, and therefore pClamp reverted into demo mode. If that's the case, you should be able to see 'demo mode' on the top left of your pclamp window, instead of the name of your protocol (if this it, you will have to recalibrate it). The last common thing to check is that your test pulse is correctly parametered (configure, seal test).
Hope it helps
Could be many things. Maybe to help others with the issue, send a snapshot of what you get on the pClamp graph (or what you get on the oscilloscope).
when i have these type of problema and chek the calibration. Used the automatic for calibrate digitazer. And maybe and not crazy problema you only need reescale the clamp test
cheers!!
Hi, I checked the snapshot you attached. Make sure to use "bath" mode - not "patch" mode - when your electrode is in bath but not on the cell yet. If you don't see a square-shape wave, you may try to re-chloride/re-connect the bath (ground) electrode and see if that helps.
Good luck!
That happens when you want to have your setup super clean. ;-)
First step to deal with this problem is attaching model cell and making sure that things look as in a book. I would start with "pipette in a bath" mode instead of "patch" mode. If this will not help next step is to verify calibration of your Digidata. I assume that you switched it off before taking the cables out of it. Crucial at this point is to make sure that you see the pulse in you seal test. If this is not the issue you may have just a noise issue with the ground electrode.
Use the model cell.
If you can get the correct signal with model cell, you cable connection and software configuration are correct. Replace model cell with your electrode holder, place your ground, if you do not have the correct signal, that tells the bad connection in the electrode holder and ground.
Check your resistance reading using your dummy cell (the one provided by Molecular Devices).Put the switch under 10 MOhms (or 0.5 GOhms) and check that the reading matches the resistance. Also Check that your analog out (#0) is the correct signal.
If you still can't see anything...call Molecular Devices. the chip in the Digidata box could of have been burned. It happens; not often, but it does. A little electrical shock and bye bye chip :)
Hi all,
By any chance, Do you know how can I write a new protocol to apply -80mV for 3 secs and 60mV for 2s and -100mV to +50mV in a 10 mV step for 10 secs?
Hi Marco,
a lot of the time passed since this question and this make the answer more difficult. Anyway, I'm trying to recall from my hippocampus and, if i remeber correctly, I checked for the reference electrode. Do you have the same problem?
Let me know.
Cheers
Luca
Hi,
I also lost the membrane test window, and i solved it by cleaning the registry of the software, see this link: http://mdc.custhelp.com/app/answers/detail/a_id/17007/~/pclamp%3A-getting-a-fresh-start-after-a-crash
After doing this, I needed to configure the Telegraph again.
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