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X8 versus DN3

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nkhanbham

Master

Posts: 53

Joined: Wed Feb 25, 2015 3:03 pm

Post Mon Feb 26, 2018 6:41 pm

X8 versus DN3

I would like to ask if people in the field have tried both of the polymers Fluidigm sell and could share their experiences? Has anyone found that switching to DN3 has given better conjugation results (ie better staining) when X8 hasnt worked well?
If there is a thread on this already please redirect.
Thanks,
Naeem
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mleipold

Guru

Posts: 5792

Joined: Fri Nov 01, 2013 5:30 pm

Location: Stanford HIMC, CA, USA

Post Mon Feb 26, 2018 7:36 pm

Re: X8 versus DN3

Hi Naeem,

I only did some head-to-head testing with a couple fairly bright markers early on (I think CD3 and HLADR were two of them).

As a general rule, DN3 gave slightly brighter signal, but also often slightly higher background, compared to X8. I don't recall the difference being more than 2x: obviously, a two-fold change from Dual=10 to Dual=20 can make a big difference in your gating, whereas a change from 200-400 probably wouldn't. And as I said, at the low end, you have to take into account whether the potentially increased background would be a problem resolving your pos and neg populations.

For my regular standard phenotyping panel, I've stayed with DN3 because it was developed with DN3. But most people use X8 with no problem, and anything from Fluidigm is going to be X8 or DM10 (which they don't sell).


Mike
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jlederer

Participant

Posts: 10

Joined: Thu Feb 08, 2018 2:42 pm

Post Mon Feb 26, 2018 9:31 pm

Re: X8 versus DN3

Hi Mike. What is DM10? Shortly after we staring working with cytof, Dvs stopped selling dn3. But I never heard of dm10. If it is dtpa it will not necessarily be better for temperamental metals. Thanks. Jim
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mleipold

Guru

Posts: 5792

Joined: Fri Nov 01, 2013 5:30 pm

Location: Stanford HIMC, CA, USA

Post Mon Feb 26, 2018 9:41 pm

Re: X8 versus DN3

DM10 is a polymer that Fluidigm made that it uses on some of its in-house commercial conjugates. They do not sell DM10 labeling kits. It's related to X8, and as far as I know is DTPA-based.

Fluidigm still does sell DN3 labeling kits, though they're not listed on their website (similar to how they also sell 25 test vials of most of their conjugates, even though they're not listed on the website). For more information, contact your FAS or local Sales associate; they can help you get a DN3 order if you want to comapre with X8.
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nkhanbham

Master

Posts: 53

Joined: Wed Feb 25, 2015 3:03 pm

Post Wed Feb 28, 2018 3:37 pm

Re: X8 versus DN3

Thanks Mike. Very helpful feedback as usual.

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