Hi–I am using a Leica SP8 confocal to image fixed immunostained tissue, and am having a problem where black horizontal stripes appear when I view samples live, and when I image them.
They appear in all of my fluorescence channels and at many z-positions, and in live view they seem to fade in and out so that sometimes the sample looks normal. Others have had this problem occasionally before on our scope, and closing and restarting LAS X fixed it in the past, but neither restarting the software alone nor turning off the microscope, scanner, lamp etc. and trying again a few hours later is fixing the problem for me now. I also have had this happen before in the first slice of a z-stack but not in the others, whereas now it is happening in many slices. Any advice on what this is or how to fix it is much appreciated!
Hi, we’ve had similar problems. Two things worked for us. First, you can try put your cell phone far away from the scope. Second, you can slow down the scanning speed. We had one user that uses scanning speed at 600HZ，even if he does not bring a cell phone with him, he still see stripes. After lowering scanning to no more than 400hz at live, then 100-200hz while acquiring, the problem disappeared. So, maybe you can try both. Good luck!
Does the striping occur even with single-direction scanning (bidirectional OFF)?
Do you see the lines if you switch from z-galvo to z-wide in the focus mode? What if you remove the galvo-stage adapter and just use a basic stage insert?
Thank you all for your suggestions–it turns out that one of the corners of our air table was not floating properly, and must have been vibrating. Increasing the pressure in that corner of the table so that it floated again made the lines disappear.