FACSCalibur - O Ring resolved, failing FACSComp now, was bubble

So, I forgot to update the blog about the O Ring issue.
Basically, we could get it to work with the O ring that was replaced by Straub, but tubes kept falling off and it just didn't feel right. The rep called to follow up and I told him what we were experiencing and that because it was the one that Straub placed on the cytometer, I felt they should send us a new one. They sent it right away and I received it the next day. It still feels a little different but is working overall, so we're happy. The rep, Oscar I think, called to check on things, which was nice.

The latest problem was that it was failing FACSComp. Torey Browning came to use the scope on Wednesday and couldn't get it to pass. She left me a message and I checked things out again. It was failing time delay, and the status showed low voltage. I reset the small black buttons, because there was a red and orange light. It didn't help. I did a regular prime and it didn't help much. Then, I did a hard prime and that worked. It must have been a bubble. Passed FACSComp.

Then, I finally got a call back from BD, Ref # 889248. The rep, who I've never spoken with before, wanted to be sure that all was well and that the problem was more than just temporarily fixed. I told him what I had done and that the sample voltage to go from 8.30 to 10.23, and he said that sounded good. He had me created a folder on the desktop called Time Delay, with two files inside: a Time Delay Cell Quest Pro document and a Time Delay Settings file to keep the settings that worked. He said our red laser looked good because the voltage was 684 (up to 750 is good, above 850 means you need a new laser). He also explained that the colored LEDs inside the machine relate to specific requirements. The event rate needs to be >200/sec, the FSC and FL4 need to be over channel 400, and the threshold needs to be above 200. Otherwise, it turns yellow. There's a window into which the events need to fall for the red light to not be on, and when the red light comes on, you're "out of range" and fail FACSComp. He told me that the lower black button is the update button and updates the range to match the events (to a certain extent) and the upper button should rarely be used because it's the resent button and removes the window altogether.

He also said that it's normal to get bubbles, as we do, and that probably they are usually in the sheath filter. To check this next time, I can bypass the sheath filter and see the effect. I had thought our bubbles were in the flow cell. We'll have to see.