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Low Tb counts with Auto tuning

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robem01

Participant

Posts: 12

Joined: Wed Apr 23, 2014 5:18 pm

Post Wed Dec 31, 2014 4:48 pm

Low Tb counts with Auto tuning

HI all,
We have recently been trying to figure out an issue with auto tuning our CyTOF2. What we are seeing is a low Tb dual counts, with reasonable oxide ratios typically less than 2%, good resolution (450 or 412), typical gas flow rates, cone currents between 3-4.5 after a deep clean of all parts. We have replaced the nebulizer, sampler cone, sample capillary line, one at a time, with new parts and have not found any to be helpful. Detector voltage relatively stable but creeping up as normal. We started using a new bottle of tuning solution around the time, +/- a few days, this pattern started.
Any thoughts or suggestions would be great to hear!!

Emily
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AdeebR

Grand master

Posts: 169

Joined: Thu Mar 13, 2014 5:58 pm

Location: NYC

Post Wed Dec 31, 2014 5:47 pm

Re: Low Tb counts with Auto tuning

Hi Emily,

How much lower are your Tb counts than they used to be? Do the Tb counts in your autotuning results match what you see if you load a fresh loop of tuning solution and manually run and review in the masses window?

There have been a couple of occasions where I've found that replacing the sample loops has given me a significant boost in tuning signals so that's something else you might want to try.

Adeeb
Adeeb Rahman
Icahn School of Medicine at Mount Sinai, NYC
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anitamkant

Master

Posts: 51

Joined: Mon Nov 18, 2013 6:30 am

Post Wed Dec 31, 2014 6:08 pm

Re: Low Tb counts with Auto tuning

Hi Emily,
Please proceed with the EQ beads sensitivity test as per the manual. Contact Tech support with the test results. They will guide you through further steps.

Regards,
Anita
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robem01

Participant

Posts: 12

Joined: Wed Apr 23, 2014 5:18 pm

Post Wed Dec 31, 2014 10:12 pm

Re: Low Tb counts with Auto tuning

Thanks Adeeb and Anita,
We are seeing around 400K to 500K tb dual counts where we typically find ~900K after a deep cleaning. Interestingly I did not see the same Tb dual counts in mass per reading window after auto tuning. 400K by autotuning and with those settings ~550 to 600K in the mass per readings window.
Anita, that was one of our next steps to check the EQ bead sensitivity and regroup Friday with tech support.
-Emily

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