Monday, October 29, 2007
FACSCalibur - Failed time delay again
I went to run FACSComp again today and again it failed time delay (signal out of range). I hit the reset buttons and did a hard prime without fixing the problem. The error changed to the "more than 5 channels difference" message. The sample voltage had been fine. So, I went to the Time Delay Update folder and got the previous settings back. Then, it was ok.
Friday, October 26, 2007
Confocal - "Line-iness" issue, in timelapse
I got a call from Austin a few days ago saying that there seemed to be some sort of "line-iness" problem with the confocal. I went up to investigate and saw that something seeemed a little weird with how their time series images looked, generally when you went from image to image. It seemed like maybe it could be due to a very subtle laser flicker, so that individual rows of pixels were slightly brighter or fainter than the ones in the same spot on the next image. Since they use pseudocolor to detect very subtle changes in fluorescence, it is more noticeable than it would be if they were not using pseudocolor.
I called Colleen who said she would make sure Steve Fryer stopped by when he was in town.
He stopped by on Thursday to check things out and he's not sure anything can be done. It was difficult to fully replicate the problem because Jason Weick didn't save the files in .ids format, but he got a sense of it. I think he thought that because we're talking pseudocolor, which is so sensitive, very subtle laser fluctuations can result in this effect. He mentioned another potential problem that is probably not the cause but would be of concern if it was. I can't recall what it was, but something like the laser stabilizer or something like that.
Meanwhile, we set up for him to come clean the system thoroughly once a year, piggy-backed onto some scopes at the vet school that are cleaned annually and will be serviced within the next few weeks. Steve Fryer emailed me to ask how the week of November 5 looked for coming out and I said it was wide open. Hopefully, we'll draw up an agreement that we'll be serviced annually and save money on travel expenses by doing it at the same time as other campus scopes.
I called Colleen who said she would make sure Steve Fryer stopped by when he was in town.
He stopped by on Thursday to check things out and he's not sure anything can be done. It was difficult to fully replicate the problem because Jason Weick didn't save the files in .ids format, but he got a sense of it. I think he thought that because we're talking pseudocolor, which is so sensitive, very subtle laser fluctuations can result in this effect. He mentioned another potential problem that is probably not the cause but would be of concern if it was. I can't recall what it was, but something like the laser stabilizer or something like that.
Meanwhile, we set up for him to come clean the system thoroughly once a year, piggy-backed onto some scopes at the vet school that are cleaned annually and will be serviced within the next few weeks. Steve Fryer emailed me to ask how the week of November 5 looked for coming out and I said it was wide open. Hopefully, we'll draw up an agreement that we'll be serviced annually and save money on travel expenses by doing it at the same time as other campus scopes.
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.
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.
Thursday, October 11, 2007
FACSCalibur - O ring not forming seal
Last night, I received an email from Jason Meyer indicating that he had problems getting the RUN light to turn green, and that the seal with the tube didn't seem tight. He also said that he changed the sheath fluid but couldn't stop it from leaking.
I came in this morning and fixed the new sheath fluid cap and checked the system. There were a few air bubbles in the line, but everything else looked fine. I pressed RUN and the light stayed amber. I changed the tube and could feel that the seal was not tight. I took apart the assembly and put it back together with no improvement. So, I called BD support. Reference number was 886802; the rep said someone would call me.
Background: Periodic Maintenance was done by Dave Straub on the 4th. Prior to the PM, the cytometer was failing FSC in FACSComp, due to high minimum scatter values. Straub's reports show it passing with flying colors after the PM. At the time of the PM, I realized that our unlabeled and PE beads had expired over a month ago. We used them anyway and I asked Eric to order some new sets of beads as well as some FACSRinse and FACSClean solutions.
10:30 am: finally received a call back. Initially, they said that consumables such as O-rings are the customer's responsibility. After telling the rep that PM was just done and that the problem occurred right after PM, he said that Straub would have replaced the O ring for us, and that it may have been tighter than expected, such that the next user might have been too forceful and bumped it out of position, causing the problem. I again took the assembly apart and reassembled it and moved the O ring around, and while I was on hold, it began working properly again. I told Jason Meyer about it, and he will be trying to run his samples soon.
I came in this morning and fixed the new sheath fluid cap and checked the system. There were a few air bubbles in the line, but everything else looked fine. I pressed RUN and the light stayed amber. I changed the tube and could feel that the seal was not tight. I took apart the assembly and put it back together with no improvement. So, I called BD support. Reference number was 886802; the rep said someone would call me.
Background: Periodic Maintenance was done by Dave Straub on the 4th. Prior to the PM, the cytometer was failing FSC in FACSComp, due to high minimum scatter values. Straub's reports show it passing with flying colors after the PM. At the time of the PM, I realized that our unlabeled and PE beads had expired over a month ago. We used them anyway and I asked Eric to order some new sets of beads as well as some FACSRinse and FACSClean solutions.
10:30 am: finally received a call back. Initially, they said that consumables such as O-rings are the customer's responsibility. After telling the rep that PM was just done and that the problem occurred right after PM, he said that Straub would have replaced the O ring for us, and that it may have been tighter than expected, such that the next user might have been too forceful and bumped it out of position, causing the problem. I again took the assembly apart and reassembled it and moved the O ring around, and while I was on hold, it began working properly again. I told Jason Meyer about it, and he will be trying to run his samples soon.
Blog start
Here is the start of the CMN Core Blog. Inside will reside records of issues, concerns, problems, and anything else that comes up with any of the equipment or services.
Each entry will show the equipment name and a brief summary about the entry in the title, and the details in the body. IT issues will have IT next to the equipment name in the title.
Anything that came up before the creation of this blog may at some point be listed in an entry, to provide background information.
Each entry will show the equipment name and a brief summary about the entry in the title, and the details in the body. IT issues will have IT next to the equipment name in the title.
Anything that came up before the creation of this blog may at some point be listed in an entry, to provide background information.
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