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Commercial cell standards?

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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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mleipold

Guru

Posts: 5796

Joined: Fri Nov 01, 2013 5:30 pm

Location: Stanford HIMC, CA, USA

Post Fri Aug 26, 2016 3:48 pm

Commercial cell standards?

Hi all,

What has been your result with using commercial cell standards in CyTOF?

At CYTO 2016, Biolegend was promoting their new Veri-cell lyophilized PBMCs. They had a poster on it as well, and their example data looked good.

So, I tried my live PBMC surface phenotyping panel on Biolegend Veri-cells and Beckman-Coulter Cyto-trols, along with a bunch of my standard control PBMCs. Note: the commercial standards are all pre-fixed in some fashion, while the control PBMCs were stained as thawed live cells. I have attached PDF layouts from FlowJo for comparison (one live control vs both commercial controls).

Overall, while running, the Biolegend Veri-cells seemed better than Beckman-Coulter Cyto-trols: less background, somewhat better staining even on some more sensitive markers. However, since they were fixed, of course there were still some differences compared to the live control.


One thing that puzzled me about BIolegend's poster at CYTO was that they showed phospho unstim/stim results. By email, Biolegend has told me that you can get custom Veri-cells made, where you could get an unstim and a stim of your choice (LPS, PMA, etc). Since they're fixed, this is probably most applicable to phosphoflow standards. And one caveat is that, unlike a live control which you stim and fix alongside your other samples, Veri-cells wouldn't be a control for whether your stim/fix worked. But as a commercial reference (even just as a way to consistently validate your antibodies), I think they could be useful.


Mike
Attachments
081116-Mike-Commercial-Beckman Coulter Cytotrol-Fixed-Raw.pdf
(1.48 MiB) Downloaded 376 times
081116-Mike-Commercial-Biolegend Vericells-Fixed-Raw.pdf
(1.56 MiB) Downloaded 436 times
081116-Mike-HIMC controls-885d5 live-Raw.pdf
(1.62 MiB) Downloaded 368 times
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AdeebR

Grand master

Posts: 169

Joined: Thu Mar 13, 2014 5:58 pm

Location: NYC

Post Fri Aug 26, 2016 8:50 pm

Re: Commercial cell standards?

Hi Mike,

We've been doing a lot of testing with Biolegend's Vericells and our experiences have been similar to yours - generally pretty good staining with the exception of some fixation sensitive markers.

I also agree that the stimulated controls are useful as a benchmark of what a successful stim should look like (though not necessarily whether your stim/fix worked). We've already been using a similar approach with in-house stimulated fixed and cryopreserved cells.

Adeeb
Attachments
VeriCell_CyTOF_1.pptx
(1008.04 KiB) Downloaded 420 times
Adeeb Rahman
Icahn School of Medicine at Mount Sinai, NYC
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mleipold

Guru

Posts: 5796

Joined: Fri Nov 01, 2013 5:30 pm

Location: Stanford HIMC, CA, USA

Post Fri Aug 26, 2016 9:11 pm

Re: Commercial cell standards?

Hi Adeeb,

Thanks for the PPT: it's nice to see that our data is pretty consistent!

I wonder how many people would be interested in Biolegend selling Unstim/IFNa/LPS/PMA+I/etc cells for phospho reference. Heck, that could be a great spike-in control sample for the CLEAN algorithm Jeff Hokanson from MD Anderson talked about at CYTO 2016.....


Mike
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billog

Participant

Posts: 15

Joined: Wed Sep 17, 2014 10:53 pm

Post Mon Aug 29, 2016 3:43 pm

Re: Commercial cell standards?

I just started looking at Vera cells, generally looks good but I don't believe out human Foxp3 stain worked. We are using Ebioscience clones and buffer system although Biolegend does claim Foxp3 works via Vera cells with their mAb and buffers I believe...

If you guys are using Ebio reagents w/ Vera cells do you also believe that EBio approach for Foxp3 is not compatible with Veracells?

Was it with cyto-trols?

Thanks, - Bill
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mleipold

Guru

Posts: 5796

Joined: Fri Nov 01, 2013 5:30 pm

Location: Stanford HIMC, CA, USA

Post Wed Aug 31, 2016 9:14 pm

Re: Commercial cell standards?

Hi Bill and Adeeb,

Bill: regarding Foxp3, I haven't tried the Veri-cells. You'd have to talk with Biolegend about how they justify their Foxp3 claim.

Adeeb: I've been talking a bit with my Biolegend rep, and she's been asking Production about custom Veri-cells. For something like a reference standard (so you don't have to stim and fix every time you make a new prep of, say, pSTAT3), she said "The product developer said that the price for phospho controls will be approximately $8-$10 at a 100 test level and ~$4 to $5 at the 500 test level."

So, $800-1000 *per stim* for 100 tests of a given stim. So, 100 tests of IFNa-stim would be $800-1000. Of course it gets cheaper the more tests you buy.

As a reference: the regular Veri-cells (so, unstim) are #425002 100 tests (4 vials) $450.00. So, stims are roughly twice the price at the 100-test level.

Depends on your volume of usage whether this is cost-effective for you, but that's the info I have so far.


Mike

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