Does anyone know if it is possible to sort micro-beads, ideally magnetic, using FACS and if so what diameters can be sorted. I was thinking of using fluorescent beads that might help facilitate this but I have no previous experience with FACS. Ultimately I want to separate beads into separate wells that can be used for PCR templates. It would be helpful if anyone knows any studies that have used similar methods. Cheers!

  • $\begingroup$ Just some questions to facilitate a good answer: What sorter do you have access to? Are they the MACS microbeads (Miltenyi GmbH)? What's bound to the beads? $\endgroup$ – CKM Jul 14 '17 at 18:32
  • $\begingroup$ I'm not sure what model the FACS is that I have access to, I can find out on Monday. We haven't decided what beads to use for our experiment yet, this is what I am trying to prepare. We want to isolate Bacteria and Archaea cells from environmental sediments but haven't decided the best way to do this yet either and what to use to bind the cells to the beads. These are all things that I'm currently considering but initially, most crucially, if it is not possible to sort beads using FACS then I will need to look for an alternative approach. $\endgroup$ – wl284 Jul 15 '17 at 10:16
  • $\begingroup$ You can sort on a fluorescent signal, though I'd be apprehensive about sorting magnetic beads. The MACS microbeads are 50nm to start, which is probably below detection for sorters not sporting a 405nm small-particle detector, making a gating strategy using scatter difficult. Have you considered a negative magnetic sort where your flow-through is your population of interest, followed by FACS? $\endgroup$ – CKM Jul 17 '17 at 18:39
  • $\begingroup$ The reason I wanted to use magnetic beads is to isolate them easily from an environmental sediment sample, not to use MACS. Therefore I could use beads larger than 50nm, I've seen some available that are several microns in diameter. The beads would just be suspended in some buffer like PBS (or whatever is suitable for FACS). Ultimately I want to then separate these beads so that each is in its own well on a well-plate, I thought maybe FACS could do this? $\endgroup$ – wl284 Jul 17 '17 at 20:05

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