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?