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Smarttube and Maxpar Direct

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Ezequiel

Participant

Posts: 5

Joined: Tue Feb 09, 2021 8:33 pm

Post Wed Feb 10, 2021 1:23 am

Smarttube and Maxpar Direct

Hello! I was reaching out to the CyTOF experts out there to get some advice with an issue. We had to collect blood samples from patients in a remote site with very little sample processing capabilities, so we decided to fix the whole blood with the Smarttube. Additionally, we have available in my lab some MAxparDirect kits that we would like to use with those samples. Unfortunately per this paper ( https://doi.org/10.1101/2020.06.26.20141341 ) looks like some of the clones in the kit are not capable of recognizing fixated antigens. Some of them of particular importance like CCR7. Did anybody encounter a way around this issue? I was thinking of trying other clones that might recognize fixated CCR7 and add them to the Maxpar mixture, but I am not sure if there are any. Any information would be greatly appreciated!!
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dtelad11

Master

Posts: 129

Joined: Mon Oct 31, 2016 6:26 pm

Post Wed Feb 10, 2021 5:40 am

Re: Smarttube and Maxpar Direct

Adeeb Rahman partially covers this in our webinar:

https://www.youtube.com/watch?v=cKNvxQciS_4

As you said, I would look into other CCR7 clones. Or rely on CD27 for central/effector memory T cells.
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Ezequiel

Participant

Posts: 5

Joined: Tue Feb 09, 2021 8:33 pm

Post Wed Feb 10, 2021 3:57 pm

Re: Smarttube and Maxpar Direct

Thank you, very interesting discussion in the video. I wonder if the "cross-reactivity" with the CD45RA and CD19 is also observed when the blood is first fixated with Smarttube. :o
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Ezequiel

Participant

Posts: 5

Joined: Tue Feb 09, 2021 8:33 pm

Post Tue Feb 16, 2021 6:30 pm

Re: Smarttube and Maxpar Direct

I would like to include here the following published protocol, as it proposes solutions for some of the issues discussed in this thread.

Kowli, S. and Maecker, H. T. (2020). Immunophenotyping and Intracellular Staining of Fixed Whole Blood for Mass Cytometry (CyTOF). Bio-101: e5004. DOI: 10.21769/BioProtoc.5004.

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