DA shutter won't open in sequence mode

I’ve recently set up our scope to do fast z-stacks in sequence mode, but I ran into a problem with the laser shutter during MDA. Our laser (an 89north LDI 7-line laser) shutters are controlled through a National Instrument PCI-6723 board, and we have a separate transmitted light shutter (a LUDL shutter) controlled through its own controller. We are using the NIMultiAnalog adapter to drive the NI board, which controls the 7 laser shutters, laser powers, and the piezo z stage (a PI P-736 stage), and receives TTL out from the camera (a Photometrics Prime BSI).

If we leave auto shutter on before multi-channel and z-stack imaging in MDA, the laser shutters won’t open but the transmitted light shutter opens properly. This happens regardless of whether we check “keep shutter open” in z-stacks in MDA. We can get around this by unchecking auto shutter and open the shutter in the main micromanager window (for any channel), then MDA would open/close the correct shutters during multi-channel and z-stack imaging. If we turn off sequence mode in the NIMultiAnalog setting, the laser shutters also work properly. If we turn on sequence mode in the NIMultiAnalog setting, but force the laser shutters to not use sequence mode (we don’t really mind keeping the laser on throughout the z-stack), we still have the same problem.

Any idea how we can fix this?
Thanks!

Hi @chenxy87,

Can you send a Trouble Report (Help > Report a Problem)? That may shed some light on the situation.

Best,

Nico

Hi Nico,
Thanks for the reply. I’ve tried Problem report but got a connection timed out error. I’ve copied the automatically generated report though. Is there a way to send this manually?
Thanks!

Xiaoyin

Either send it to my personal email or use the upload function here.

Hi Nico,
Thanks. Please see the problem report in the attachment.

Xiaoyin
ProblemReport_XC.txt (367.2 KB)

I think (but not absolutely certain) that this is caused by having the AO voltages in your channel1 definitions. Those are basically a second way to shutter your lasers, and certainly will lead to confusion.

Taking those out of the channel1 definitions should help. You can either add them as one AO voltage per group (resulting in a slider in the UI), or set them all to 5 V in your System-Initialize group. B.t.w., if you rename “initialize” to “Startup”, it will automatically be applied when you start Micro-Manager.

Hi Nico,
Sorry I got swamped by other things for a while and just had a chance to revisit this problem.
I tried this and it didn’t solve the problem. I’ve attached the new problem report.
I created a new group called “ImagingChannels” that only have the filters and changes the core shutter. The problem is exactly the same as before, i.e. the shutter won’t open when in sequencing mode if the autoshutter is checked on the UI, but will work either the sequencing mode is turned off or autoshutter is unchecked and the current shutter is opened manually before imaging.

newProblemReport.txt (856.1 KB)

Hi @chenxy87,

Is there a problem leaving sequence mode off? Sequence mode allows the computer to upload a sequence of voltages to the NI board, and the NI board will cycle through those voltages (rather than listen to computer commands to set voltages). The DA Shutter has no knowledge of these sequences, hence the MDA will not upload a sequence of voltages. I think that the DA Z-stage will understand this, and would be able to run a fast Z-stack using the NIMultiAnalaog adapter. It is a bit of a pity that the NIMultiAnalog adapter does not implement “blanking”. Anyone could add that functionality, but until that point in time I would simply not switch on sequence mode in your situation.