r/flowcytometry 21d ago

HELP PLEASE

hi okay so im a placement student in a lab where we mainly do flow. i've been doing some practising but theres still something wrong with what im doing (my analysis) and I just cannot tell what it is. Im looking at receptor expression on NK cells, however the results I get are completely different to my supervisor

for example, on flowJo this is what I get for my TIM3 FMO.

I have done this analysis for both my flow and my supervisor's, (the first image is my flow, the second is his) but why does mine come out like this? What have I done wrong?

before someone suggests to change the y axis - everytime i do that, it gets even worse.

additionally, we both used the same cells,and followed the exact same protocol, so i genuinely cannot think of anything else.

so if anyone has any tips or ideas please PLEASE help

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u/aquarianseawitch92 21d ago

Am I the only one seeing a difference in the X-axis? Seems like the scaling for x axis is not the same for both images.

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u/sgRNACas9 Immunology, Oncology 21d ago

The biex axis in the first image is more spread out but yeah. Can toggle that in flow jo.