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Stain cells before barcoding

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MarcelaAH

Participant

Posts: 6

Joined: Fri Jan 16, 2015 10:20 pm

Post Wed Jan 18, 2017 3:50 am

Stain cells before barcoding

Hi all

I am considering using the MaxPar barcoding kit from Fluidigm. However my stain is not optimal after fixation. Does anybody know if you can stain your cells separately first and then barcode and mix? It would require more antibody but the acquisition would be more uniform. Any comments or experiences are greatly appreciated.

Marcela
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mleipold

Guru

Posts: 5792

Joined: Fri Nov 01, 2013 5:30 pm

Location: Stanford HIMC, CA, USA

Post Wed Jan 18, 2017 3:56 pm

Re: Stain cells before barcoding

Hi Marcela,

Yes, you can do surface stain of live cells, then fix, then do Fluidigm barcoding. I've heard of at least two different groups doing it.

It will mean that there will be variation in surface staining and you won't save as much on reagents, but if you're doing intracellular staining, it should be more uniform.


Alternatively, you could do the CD45-barcoding as in the Mei et al papers; that was built for barcoding live cells.


Mike
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DKaufman

Participant

Posts: 3

Joined: Wed Jan 18, 2017 9:02 pm

Post Fri Mar 24, 2017 7:40 pm

Re: Stain cells before barcoding

I am wondering if anyone who has a CyTOF 1, has purchased and successfully hooked up a Super Sampler. I have followed the instructions that came with the sampler to download the software and connect the tubing and serial cable. But I think I may not have the CyTOF on the correct settings. The instructions are biased toward the Helios and CyTOF 2 and so when it shows the settings under the Devices tab for the Syringe Pump for CyTOF 2, the drop down should be empty and set to COM5.

Once I close the CyTOF software page, configure the device through the Super Sampler software and push initialize, I continually get a error message saying the port 5 is not available. Then another error message comes up that says "Unhandled exception has occurred in your application." I have tried multiple other ports with the same error.

Any thoughts or suggestions would be greatly appreciated!

Thanks,
Dawn
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BjornZ

Contributor

Posts: 43

Joined: Fri Jul 10, 2015 1:04 am

Post Fri Mar 24, 2017 10:19 pm

Re: Stain cells before barcoding

Hi Dawn - Can you please send me an email (zbbjornson at gmail.com or info at victorianairships.com)? Will get you up and running right away.
Thanks,
Zach

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