QC for 120um and 150um nozzles

[kdomenico 0]

I am new to the Astrios and to MUG. I will not be using the standard nozzles for sorting on my Astrios. I will be using the 120um and 150um nozzles. I have a question regarding QC. I was able to pass QC with the 120um nozzle once, but haven't been able to since. I have never been able to pass QC with the 150um nozzle. If these nozzles will not pass the QC program, what is the best approach to determining if the Astrios is in alignment and OK to sort with? The only idea I have come up with is to use the 100um QC for a target - meaning if the bead peaks from a larger nozzle hit the "target" peak channels off the 100um nozzle than I'm good to go. Any other ideas and suggestions are welcome. Thank you!

[jamesmccracken 0]

Hi Karen,

 

You should still be able to pass QC on either of these nozzles, as Alignment QC is based on being able to align to the core stream and the width of the sheath stream won't effect that.

 

I'd make sure you have really gotten all the air out, especially from the nozzle assembly. Vertical stream is important, as is beads at a good concentration to allow for a low pressure differential when measuring CVs.

 

If you still can't get all parameters to pass, think about what you are sorting and if those parameters will even be needed. If your far-red channels fail, but you are doing a violet channel only sort, then no problem.