Flow Cytometry Controls (Intro to Flow - Episode 5)

Flow cytometry is a powerful technique for studying single cells. In this episode of our Introduction to Flow Cytometry series, I dive into the controls you will need for a flow cytometry experiment. This includes compensation controls, FMO controls, and isotype controls. Flow cytometry loves controls!
Intro to Flow Cytometry Playlist: • Introduction to Flow C...
Slide deck: drive.google.com/file/d/1zsKI...

[@yasminenonose2597]

This video is a public service! Thank you so much!

[@nataliafedianina3808]

Great and simple overview of controls. Thank you

[@ajarieger_flow]

Thanks!

[@nguyen-anhnguyen3325]

Thank you so much. It's such a great overview and introductory to controls!

[@ajarieger_flow]

Thank you!

[@emberthong7503]

What a useful video!! Thanks :)

[@ajarieger_flow]

Glad it was helpful!

[@jaimezaldivar189]

Awesome. With you flow cytometry is fun

[@ajarieger_flow]

Thank you ☺️

[@TheCheerup123]

thank you so much! its really helpful for an introduction to flow!

[@mojtabazareie9382]

What a useful video! Thank you Aja 🙏🙏

[@rosealuochojuok9898]

This video has been so useful to me...thanks

[@tunaonal7496]

Thank you for your great video! 🙏🏻

[@ajarieger_flow]

Thanks! 😊 Stay tuned- I’ll have more coming in the new year!

[@yangwang6805]

Your videos are so helpful, one of the best FC videos I can find on youtube. I promise nobody watches your videos more careful than me, I learned word by word. Just curious, why this video can not turn on the captions? Other videos can turn on captions, thank you

[@ajarieger_flow]

Thanks for the comment! I honestly didn't think of adding them- but am working through my videos to add them in and will have them in all future videos :)

[@yangwang6805]

@@ajarieger_flow Hi Aja, thank you so much for your reply. I am actually learning to analyze flow cytometry data recently. Do you have any plans to make some videos to interpret the FC output plots? Like histogram, density plots, all how to gate the margins? Such like this, I tried to understand them first then will have a better understanding of the data analysis. Thank you again for the wonderful videos you made, you helped me a lot for my new job! ❤️

[@vanessateixeira1809]

Excellent video series!!! Extremely educative!
I'd like to ask you two questions I often see professionals struggling about:
1 - Considering this 4 colors panel example, In your opinion, is it necessary to perform all 12 control tubes in all the the experiments performed? (e.g. patient samples will be collected over time to form groups, so the same experiment will be performed multiple times with different samples). Which ones are a "must have" in all the experiments, besides the PC and NC?
2 - Can we use beads for compensation even though the experimental samples will be cells?
Thank you in advance.

[@ajarieger_flow]

Thanks for reaching out- I'm glad it was helpful for you. (1) My recommendation is always to start with all necessary controls and then determine what is needed for your particular experiment. For example, you may discover that certain markers don't need an FMO or that you need patient-specific FMOs for certain markers and not others etc. etc. etc. There are so many potential iterations here that the answer is the always flow answer of "it depends". But always start by testing them all for yourself and determine this empirically. (2) Yes using beads is fully acceptable. Just make sure they follow the same rules for compensation controls as if you were using cells.

[@vanessateixeira1809]

@@ajarieger_flow Thank you so much for all the replies! I'm binging-watching all your series !! It has been very helpful in both reviewing the why for many practices we learn from other people but don`t really know why, and also getting some extra tips.

[@happyeagle92]

How do you actually create an isotype control? Do you just use random other markers that have the same isotype as the antibody? Or would I need to buy a specific isotype antibody to stain my cells with?

[@ajarieger_flow]

You will need to buy a matched isotype control. You want it to be the same type of antibody (i.e. IgG) and have the same type of light chain. If you check out our video on selecting antibodies, we go into this there :)

[@gvil1313]

Hello! How can I tell if my cells have a lot of Fc receptors?

[@ajarieger_flow]

There is pretty good information in the literature for most cells. I would start there. And if I’m doubt, it doesn’t hurt to block!

[@mannyapp1214]

Can explain unstained

[@ajarieger_flow]

Unstained cells are cells that you have added only buffers to- no stains of any kind.