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New WB injector and cell acquisition solution

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It is fine to promote your company's reagents. Just make sure they are relevant to CyTOF, and do so in moderation and style :-)
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kunicki

Contributor

Posts: 38

Joined: Thu Apr 13, 2017 8:46 pm

Post Wed Aug 15, 2018 9:50 pm

Re: New WB injector and cell acquisition solution

Hi Mike,

Do you mind explaining how you calculated your CV values for all/each of the slides?

Best,
Matthew

mleipold wrote:You can find a PDF of our results here (file was too big to attach here)....let me know if you have trouble accessing it: https://drive.google.com/open?id=1LxvF5 ... IZUqbywInQ
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mleipold

Guru

Posts: 5796

Joined: Fri Nov 01, 2013 5:30 pm

Location: Stanford HIMC, CA, USA

Post Wed Aug 15, 2018 10:11 pm

Re: New WB injector and cell acquisition solution

Hi Matthew,

For both the Medians and the CVs, I gated to the indicated population in FlowJo, then used FlowJo to calculate Median and CV for the specified channel.

For example, I think two of the entries was "CD4+ CD4 Median" and "CD4+ CD4 CV".

In those cases, I gated down to CD4+ (Live Intact Singlet/CD14- CD33-/CD3+/CD4+ CD8-), then had FlowJo calculate the Median and CV of the CD4 signal for that population.


Mike
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kunicki

Contributor

Posts: 38

Joined: Thu Apr 13, 2017 8:46 pm

Post Wed Aug 15, 2018 11:25 pm

Re: New WB injector and cell acquisition solution

Thank you Mike,

So this isn't the CV% for CD4+ T cell frequency between replicates, right?

Did you see any differences in the CV% between replicates using the two methods? What was the range of CV% you observed across populations?

Matthew
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mleipold

Guru

Posts: 5796

Joined: Fri Nov 01, 2013 5:30 pm

Location: Stanford HIMC, CA, USA

Post Wed Aug 15, 2018 11:35 pm

Re: New WB injector and cell acquisition solution

Correct, all those values are Median and then the CV of the signal intensity. There is no Freq data Median or Freq CV in that presentation.


There were no independent replicates in that experiment: it was stained all the way through Ir, then split into thirds, and run under each condition (NB+MilliQ, WB+MilliQ, and WB+CAS).

I didn't notice any obvious difference in Freq between the 3 replicates in the surface phenotyping. We're in the middle of some additional testing that might allow me to answer that question. So far, everything's been resolved and gate-able, so Freq hasn't really been an issue.


Mike
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AdeebR

Grand master

Posts: 169

Joined: Thu Mar 13, 2014 5:58 pm

Location: NYC

Post Mon Apr 08, 2019 4:45 pm

Re: New WB injector and cell acquisition solution

Hi all,

Thought I would update this thread with our recent pre-print showing the results of our testing of the WB + CAS acquisition protocol.

"A Modified Injector and Sample Acquisition Protocol Can Improve Data Quality and Reduce Inter-Instrument Variability of the Helios Mass Cytometer"
Brian H Lee, Geoffrey Kelly, Shermineh Bradford, Melanie Davila, Xinzheng V Guo, El-ad David Amir, Emily M. Thrash, Michael Solga, Joanne Lannigan, Brian Sellers, Julian Candia, John Tsang, Ruth Montgomery, Stanley Tamaki, Tara Sigdel, Minnie Sarwal, Lewis Lanier, Yuan Tian, Cheryl Kim, Denise Hinz, Bjoern Peters, Alessandro Sette, Adeeb Rahman
bioRxiv, posted April 05, 2019
https://doi.org/10.1101/600130
Adeeb Rahman
Icahn School of Medicine at Mount Sinai, NYC
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