Re: New WB injector and cell acquisition solution
I know there are a few places that are testing out the new wide-bore (WB) injector + CAS as recommended recently by Fluidigm, rather than the previous narrow-bore (NB) + MilliQ.
Stanford HIMC did our own testing, using all 3 of our standard PBMC assays. Basic protocols can be found below.
surface phenotyping: http://www.bio-protocol.org/e1382
ICS: http://www.bio-protocol.org/e1370
phosphoflow: http://www.bio-protocol.org/e1496
You can find a PDF of our results here (file was too big to attach here)....let me know if you have trouble accessing it: https://drive.google.com/open?id=1LxvF5 ... IZUqbywInQ
We present most of the data in a similar fashion to how Fluidigm presented their data: Median and CV ratio of (WB+CAS)/(NB+MilliQ) for several populations.
In short: in our hands, for all 3 of our assays, the previous method (NB+MilliQ) was as good as, and often *better* than, the new method (WB+CAS). For the surface phenotyping, normalization (Fluidgm or Finck) made the ratios more comparable, but never flipped it.
Since the new WB+CAS was no better than the original NB+MilliQ *and* there is also a significant increase in daily instrument maintenance due to salt buildup from the CAS, the Stanford HIMC is not planning to implement this with our current protocols.
Note: it's entirely possible that there *are* protocols where this new protocol would make improvements, like Dariush saw. However, it's definitely *NOT* every case......I'd recommend that your site do your own testing (ideally for each assay) to see if there's a difference.
Mike