FAQ  •  Register  •  Login

Maximum panel size with CyTOF

<<

siveenks

Participant

Posts: 1

Joined: Thu Jun 11, 2020 5:13 pm

Post Mon Sep 07, 2020 12:12 pm

Maximum panel size with CyTOF

Hi,
I am not a CyTOF user as of now, but we may get one later next year. I am following the discussions and publications on CyTOF for the last few months to keep myself connected to the technology and recent publications.

I have a basic doubt. The Fluidigm website says that CyTOF can work with more than 90 isotopes. But, what is the largest panel published with CyTOF till now? Mostly I can see 30-40 antibody panels and I haven't seen anything above this number. I would like to get any updates on the largest panel currently proven to work on CyTOF.
Regards,
Siveen
<<

MCOlivier

Contributor

Posts: 34

Joined: Mon Oct 05, 2015 9:48 am

Location: European Genomic Institute for Diabetes, Universitity of Lille, Institut Pasteur de Lille, France

Post Tue Sep 08, 2020 6:48 am

Re: Maximum panel size with CyTOF

Dear Siveen,

I think the bigest panel published is 42 antibodies, but I'm not sure. I am analyzing a panel with 46 antibodies, part of which were labeled with "exotic" metals, as 113In, 115In, 157Gd... Considering new Cadmium labeling kits released by Fluidigm at the end of 2019, you can easyly reach 50 antibodies (but this implies not using the barcoding kit From Fluidigm based on Cadmium Isotopes).

Hope this helps !
Best
Olivier
<<

tomash

Participant

Posts: 19

Joined: Sun Oct 19, 2014 10:15 pm

Post Tue Sep 08, 2020 9:38 am

Re: Maximum panel size with CyTOF

If you'd like a simple 'rule of thumb' of achievable panel sizes:

1. Using just the typical/easy/broadly used reagents, < 40. For example, using 'off the shelf' reagents I think you could do a panel around ~37 abs (not including barcoding etc).
2. Including non-standard (e.g. Indium 113 and 115, barium 139, bismuth 209 and Yttrium 89) and newer metal reagents (palladium series, cadmium series), often involving self conjugations, probably 40-50. This does depend on exactly which reagents you use, and how they are being used. e.g. if the palladium channels are used for barcoding, then you can't use them for actual measurement antibodies, etc.
<<

dtelad11

Master

Posts: 129

Joined: Mon Oct 31, 2016 6:26 pm

Post Tue Sep 08, 2020 12:56 pm

Re: Maximum panel size with CyTOF

The Astrolabe database has over 1,500 experiments. The largest panel is 47 markers, and it's publicly available:

https://www.sciencedirect.com/science/a ... 9120300999

Based on data from the past year, a typical experiment is 40-43 markers, plus potentially three to nine barcoding channels.
<<

mleipold

Guru

Posts: 2640

Joined: Fri Nov 01, 2013 5:30 pm

Location: Stanford HIMC, CA, USA

Post Tue Sep 08, 2020 3:31 pm

Re: Maximum panel size with CyTOF

Hi Siveen,

I think it's important to differentiate between theory, current status, and potential future practice.

The mass window of the Helios is about 76-210 (about 130 mass units). So, in *theory*, you could use up to 130 different probes.

In *practice*, some of these mass channels will never be usable for probe-labeling. For example, if you look at isotope lists (eg, http://www.chem.ualberta.ca/~massspec/a ... _abund.pdf ):

Ar-dimer: 40 x 2 = 80

Kr (gas): 83
Xe (gas): 129, 131

There will *never* be probes on those channels because they're masses occupied *solely* by gases, and/or Ar-dimer which is too high a background to ever have a probe on it.


The *current status* has probes (chelators or otherwise) for Y, Ru, Rh, Pd, Ag, Cd, In, Te, I, La, Ce, Pr, Nd, Sm, Gd, Eu, Tb, Tm, Dy, Ho, Er, Yb, Lu, Ir, Pt, Os, and Bi.

There are a couple papers which have used Au, Re, Cs, Nb, Zr, and Ta.


But, that still leaves several elements like Sn, Pb, and Mo which are in the mass window, but haven't been formed into a stable probe (yet?). As illustrated last year by the Cadmium channels, the nice thing about mass cytometry is that once you can form a stable probe with one isotope of an element, you automatically can form a stable probe with *all* isotopes of the element that exist in nature and that you can obtain at sufficiently high purity.

So, for "future practice", give the chemists time, and there will hopefully be more channels opening up (assuming they don't overlap with current channels from other elements, of course).


Mike

Return to CyTOF general discussion

Who is online

Users browsing this forum: No registered users and 1 guest