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Magnetic beads in CyTOF

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Delgonan

Participant

Posts: 2

Joined: Tue Apr 30, 2019 2:38 pm

Post Tue Apr 30, 2019 4:03 pm

Magnetic beads in CyTOF

Hi everybody!

I'd like to perform an experiment using the magnetic beads Dynabeads (3 microns in size) and I'm concerned about its feasibility. Has anyone used them? I've been reading some topics and it's not clear if someone has used them in CyTOF, not for markers enrichment, but as the matrix of the experiment.

I'm quite concerned about the magnetic beads clogging/getting stuck and the iron particles harming the detector.

Thank you!
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mleipold

Guru

Posts: 2095

Joined: Fri Nov 01, 2013 5:30 pm

Location: Stanford HIMC, CA, USA

Post Tue Apr 30, 2019 11:16 pm

Re: Magnetic beads in CyTOF

Hi Antonio,

Here's an earlier thread: viewtopic.php?f=6&t=293&p=905&hilit=dynabeads#p905

I'm not aware of anyone using them as the "cell", though.


The main issue with any of these beads isn't so much the iron (that will hit the cones, but it won't make it through the current mass range of the quadrupole filter, so it won't reach the detector), but any other impurities that may be in the beads. Lead, tin, cadmium, and a few other relevant things *are* in the mass range.


In short: you might just take some of the beads (washed a few times in buffer) and run them on the CyTOF in Event mode. And/or, dissolve them in concentrated nitric acid and run that (well-diluted!) in Liquid mode.

Either should show you if there are any unexpected impurities.


Mike
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GregBehbehani

Master

Posts: 80

Joined: Tue Apr 12, 2016 10:17 pm

Location: The Ohio State University, Columbus, Ohio

Post Wed May 01, 2019 12:57 am

Re: Magnetic beads in CyTOF

Hi Antonio,

I would agree with Mike; it's the heavy metal contaminants that will be the problem. I would follow his suggestion to dissolve them in acid and run them in solution mode starting at a 1:1,000,000 dilution and then increasing the concentration in 10-fold steps until you start to see metal. Once you know how if and or how much contaminating heavy metal there is, you could then try running the whole beads (if the solution data suggests it will be safe).

Good luck,

Greg
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Delgonan

Participant

Posts: 2

Joined: Tue Apr 30, 2019 2:38 pm

Post Wed May 01, 2019 9:46 am

Re: Magnetic beads in CyTOF

Hi Mike and Greg,

Thank you for your answers. I'll definitely follow your advice and dissolve the beads in nitric acid, dilute them and run the liquid mode.

Antonio

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