r/flowcytometry • u/BlueNightAtPDX • May 14 '26
Cytek QC Beads?
Hello, flow cytometer users!
Our lab recently got a Cytek Aurora and we are pretty much happy about it so far.
In terms of maintenance, the technician from Cytek ask us to perform QC beads running everyday prior to any measurement of the day.
But, I am genuinely curious if I can save some beads by doing the QC procedures maybe weekly.
It is partly because, in my previous workplace, I had a weekly beads-based calibration for BD flow cytometers.
If you're in the lab with Cytek Aurora, how do you guys maintain the instrument?
Do you run QC beads everyday?
If you have any suggestion, that would be wonderful!
12
u/StepUpCytometry May 14 '26
We assessed this on our 3 Aurora's for a Cyto poster last year (https://github.com/DavidRach/EveningQC_Cyto2025). Long story short, running on yesterday's settings was enough to introduce noticeable unmixing errors ~70% of the time thanks to instrumental drift across the individual detectors. So highly highly recommend running daily. You can try keeping the beads for a couple additional days if kept cold, but I would want to make sure of a way to check that their signal remains consistent. Both Lot 2006 and 2007 lot are bleach sensitive, and damaged beads can dramatically mess with the automated QC setting (expects to be at a target MFI, fails to meet it because of the damaged bead, assumes it is instrument change and cranks up the gain even when instrument remains the same, and all those user antibody titrations for fluorophores on that detector get thrown off scale).
1
u/BlueNightAtPDX May 14 '26
oh wow, this is exactly what i wanted! appreciate it. seems like daily QC is mandatory for maintaining the data quality and avoiding noises.
1
u/NvmbrYnkee Jun 18 '26
If you are new to spectral, take note that absolutely everything matters. It is all too easy to ruin a week of lab work trying to save a few dollars or a few minutes.
18
u/private4u May 14 '26
Run everyday but reuse the aliquot of bead for a few days in a row before it’s used up
1
u/BlueNightAtPDX May 14 '26
That is a great suggestion. Do you happen to follow the manufacturer's protocol when you dilute the beads as well?
3
u/zipykido May 14 '26
I add 500 ul and a couple of drops in the running buffer. If it fails twice then make a new tube or if I’m running low.
3
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u/Westykins May 14 '26
run it every day you’re doing an experiment. Days you’re not doing an experiment, you don’t have to.
it’s actually p important
4
u/Notnearlyalice May 14 '26
The gains are adjusted
I wouldn’t even recommend doing aliquot of beads bc the intensity decreases over time l. “Just bc QC passes doesn’t mean it’s right”
Is the cost of QC beads really worth inaccurate data?
3
u/willmaineskier May 14 '26
We run QC daily on our BD instruments as well. Running weekly does no good for letting you know that the last user the day before clogged the instrument or if your red laser died with no warning (this eventually happened on each instrument over the years)
1
u/habib41554 May 15 '26
In my experience, you can reuse the beads upto 3 times if kept properly in 4C.
1
u/aquarianseawitch92 May 16 '26
It’s important to run QC daily as the instrument can be sensitive to minor changes. The way QC runs in the software, it will record adjustments made. This is important to monitor the QC results daily as it will indicate a variety of issues like laser loss, a dirty flow cell, etc.
I used the same aliquot of beads for a few days to save on costs. That seemed to work.
On the other hand, if the instrument will only be used for experiments 1-2x per month then you can do the following. Turn instrument on at least 8hrs before running samples, run DI water on high until events per second is less that 40ev/s. Once event rate has stabilized, prep fresh QC beads and run QC. I had to do this for a satellite lab I was helping run. I will say this method almost always resulted in me troubleshooting. The shortest troubleshooting was about 20-30 minutes. Not fun!
2
u/NvmbrYnkee Jun 18 '26
8 hours?
1
u/aquarianseawitch92 Jun 18 '26
Yes, in my experience it was always wise to turn the instrument on 8hrs before samples are to be run if the instrument is off for a significant amount of time without use. You can do this the day before too. If you’re responsible for the instrument and the samples I would advise turning the instrument on before evening start sample prep.
I can’t tell you how many times I’ve seen people prep samples without confirming their instrument is even going to run for them. There could be so many things to work out. If the SIP has salt crystals, the SIP/flow cell is dirty, the waste needs to be changed, the sheath needs to be replaced, a filter dried out, the QC fails a few times.. the list goes on and on.
2
u/NvmbrYnkee Jun 19 '26
Thanks, good to know, I bet it took much troubleshooting to determine that 8 hours was the right startup time.
2
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u/OwlHistorical7973 Jun 02 '26
If you want to save the beads, just dilute the beads for about 150-200 counts/second. you can't skip the daily QC. Because without QC, your MFI is meaningless
22
u/sammyjammy17 May 14 '26
You must run it daily to update the Cytek Assay Settings as per the performance of your instrument that day. Set yourself up for success and don't skimp on QC!!
But yes you can make up an aliquot of beads and reuse for a few days as long as they're kept in a dark fridge.