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Issue with a particular detection channel in CyTOF

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nlazarus

Contributor

Posts: 20

Joined: Wed Nov 13, 2013 7:28 pm

Post Tue Jun 24, 2014 6:13 pm

Issue with a particular detection channel in CyTOF

Has anyone experienced an issue with variable signal detection in a particular CyTOF channel - as in sometimes you see a signal, other times you don't? This is not about a reduced signal but rather all or nothing.

Scenario: IgA-148, staining human PBMCs. The commercial DVS reagent worked well for several runs until I ran out and ordered more. IgA signal went blank for 2 runs using the new vial of IgA- even when using the same donor sample. Ordered and tested another new vial of IgA-148 (same lot)- still no signal. DVS provided a pro bono additional vial (also same lot - only 1 lot of IgA has been produced) and I also made my own in-house conjugate...tested all 4 anti-IgA-148 (May 8,2014) and the signal in 148 reappeared weakly for 2 vials and strongly for the other two. Proceeded to restain a previous "known" sample along with a new patient sample (June 5, 2014) and once again, there was no signal in 148. Same occurred on June 19, 2014. This phenomenon does not seem to correlate with a particular donor or whether the sample is fresh or frozen.

I have attached a PDF with the stainings if anyone has the interest and/or the time to review it.

There is really no way that I could have forgotten to add the IgA each time the 148 signal disappeared....unfortunately, I did not have another conjugate in 148 to run as a positive control...but I will purchase one as I move forward.

Has anyone else experienced such an issue or heard of one?

Thanks!

Nicole
Attachments
IgA Example for CyTOForum.pdf
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anitamkant

Master

Posts: 51

Joined: Mon Nov 18, 2013 6:30 am

Post Wed Jun 25, 2014 2:35 am

Re: Issue with a particular detection channel in CyTOF

Nicole, Please contact tech support. Once we solve the problem we can post reply.
Thanks
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Ofir

Master

Posts: 75

Joined: Thu Nov 07, 2013 12:46 pm

Location: US, CA

Post Wed Jul 02, 2014 7:15 pm

Re: Issue with a particular detection channel in CyTOF

Hi Nicole,
This is indeed strange. It is reminiscent of my issue of several antibodies not working in one particular experiment.
See details here: viewtopic.php?f=4&t=150

Ofir

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