Its been 7 weeks since I got back from Cytopeia and I have done little other than run Influx since I got back, the hours spent on the instrument are already too many to count and its paying off in unimaginable ways. We got over 80% efficiency in single cell sorting last week, one cell per well, with 3 from 16 cells coming up with positive bands after PCR. On repeat with the next 16 wells a further 4 came up positive. An impressive result from a sort that was set up after 5 hours battling a failed sheath pressure transducer on FACSAriaIIu only to bail once we realised the plate wasn't able to respond in the Y-direction (later to find the serial port had not been plugged back in after a sample agitation motor was replaced - but thats another story). So, 5 hours down, almost 12 hours in the day with half an hour lunch, the Influx was tackled head on with a 4 colour, 4 laser sort and did it ever shine. We had some problems with drop charge contacts and tracked the problem down using scope probes, multimeters, plate voltage read outs, and after replacing the contacts on the shoe behind the nozzle assembly a second time (this is why it was overlooked - twice was totally unexpected) we have now got some seriously reliable side streams. What I have benefited from most here is the knowledge of changing sort driver racks, trouble shooting over the phone with Brent, learning about software, getting the builds installed and updated, replacing high voltage boards etc. It has been a thorough learning experience and I can now say the setups are taking me an hour, the experiments are taking an hour to set up and the sort button is ready to hit after 2 hours, 11 colour panels, 6 way sorting. Sweet. Its been nothing less than epic but feel we are well on the way to some amazing sorts in the near future, first up being cell cycle sorting and microparticles. Also, if anyone has ideas about esophageal cell markers I would be most interested.
So thats me for April, will keep you updated as things keep progressing. Thanks BD for the opportunity to talk in Adelaide, I look forward to the next one.
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