Sunday, June 27, 2021

Eclipse + FAIMS + 20 nanoliter/minute + NanoPots!


I can't remember if I posted on a preprint of this study or if I've just read it so many times that I thought I did

How far can you push an Orbitrap Eclipse for direct single cell (no SCoPE-MS or boost or DIA) just DDA? This might be a good answer! 

One normal-sized human cell, prepared with NanoPots, ran at 20 nanoliter/minute (they used a Dionex RSLC 3000 at 250 nL/min and split the flow lower to get it to 20 nL/min). There are some really interesting comparisons here. Eclipse, so ion trap MS/MS or Orbitrap MS/MS? FAIMS on vs FAIMS off? What fill times? What AGC targets? 

The total gradient time appears to be around 150min, so not nearly as bad as I would have assumed to get to a flowrate that low. A custom 50cm column is used, I think, but I forget the details. I don't believe they used match between runs to get to their total numbers. Data processing was in Proteome Discoverer. A great reference for this field of single cell proteomics that is 

(Zoolander quote moratorium is over! I get 4 more before it kicks in again)

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