Flow Cytometer Basics and FLOWJO Analysis

This video is a crude and basic introductory lecture on how one can use FACS and analyze data on FLOWJO. The question we are trying to answer he is whether a certain tumor cell expresses Protein X? The video is intended to give you a mild flavor of what to expect before you plan a FACS experiment and also to give you a heads up on what questions to prepare before you seek assistance from a Senior scientist or a Post-Doc.
For any dumb questions or concerns on FACS or FLOWJO please contact me at : - [e-mail]
I am a novice looking forward to learn through teaching others.

[@sachingcopk]

Nice information ...great to start with flow cyto

[@gabrielaquiroz8089]

You explained it very well!! thanks a lot!

[@kalyanghatak2023]

Thank you so much for your explanation. Helpful for beginners.

[@Onyxscubababy]

Nice quick tutorial! It's funny that the "Eat a banana" reminder appears in the video:-)

[@themrvims]

Onyxscubababy Thanks for watching. Yes need the daily potassium intake!

[@rubypina41]

Thank you so much for making this video!!! It’s very clear, now I have an idea how to analyze my data in FlowJo!!

[@themrvims]

Thanks for watching! Yes, Flowjo is a great tool!

[@fatimahameedat4453]

Thank you very much, appreciated.

[@ndroo1019]

Thank you for this video, it was very helpful in learning how to use flow jo

[@themrvims]

Thank you. That’s a lovely Canine

[@shilpeechanda2233]

Well explained! Liked the way you speak and your voice 😀.

[@xkid06]

This helped me start out with flowjo, I appreciate it! And I hope you ate your banana :)

[@graviyt]

very informative video!! Thanks so much!

[@sachingcopk]

Very informative video ; nice to watch for the beginners

[@themrvims]

Thanks Sachin. I think I could’ve done a better job.

[@wuprabhu]

Thank you for the video. I have used cFlow and FACS Suite but I have never used Flow jo. its very helpful to understand how the flow jo analysis works.

[@themrvims]

Glad to help Dr. Ramamoorthy. Please let me know if you have more questions. The analysis shown on the video is very basic.

[@mahassenbarboura5633]

thanks for this tutorial , so clear

[@themrvims]

Thank you! Feel free to let me know if you need more information.

[@nahuelcesattilaluce1257]

thanks a lot! did you find flowJo for Mac somewhere for free? I had not luck with that

[@catnipwho]

The parind rises again

[@Levis050295]

Thank you for the video, really helped me out =)

[@themrvims]

Thanks for watching! I’m glad it helped you. All the best!

[@jess_travel_bee6108]

Thank you Vimal, this was helpful to get started :-) Greetings from Germany

[@themrvims]

jess_travel_bee Thank you taking time to watch the video. Yes this is just a very brief and amateur introduction to flow cytometry. Now that I have gained some more experience, I am planning to make a better and more lucid video. Feel free to connect if you need any more assistance.

[@JamesDinh-th4yr]

Such a great video!

[@themrvims]

Glad you liked it!!

[@kyweyokekalay]

I've watched your video and love it. I am a new PhD student, who need alot of help with FACS data to analyze and how to use FlowJo. I am wandering if you are free to glance at my data and help me. I want to learn from you.

[@themrvims]

Hi Kay,
Apologies for the delayed response. If the requirement still stands I can go over the data and give you some insights. My email is in the description.

[@sallygalal4544]

Hello, I am seeking professional help for data analysis and wondering if you could help with that. Thank you!

[@pedrammahmoudi4052]

Good video for introduction, but you could have cut the last part, where your data sets didnt work out.

[@themrvims]

Thank you for the feedback. Yes I chose to keep it. It was my bad that my files were not organized. But I felt it might be useful just in case. Hope it wasn’t too confusing.

[@cacevedo07]

This has been the best tutorial on FlowJo. Don't apologies for your mistakes, every time you made them we learn a lot from them. I have a couple of questions:
1) How many cells do you bring to the sorter? like, how many cells would you consider a good concentration to sort and not clog the sorter?
2) What volume of cells do you bring your cells to be sorted? Like 500,000 cells in 1 mL buffer?
3) What flow rate do you usually use to sort your cells?
Many thanks.

[@themrvims]

Thank you so much for the appreciation!
Most of my experience was in Flow Cytometry and not sorting. But a general rule of thumb would be to have a lower cell density since there is a good chance of aggregation / clumping at higher densities. Also, it's better if your events/sec do not exceed 10,000. You can try to start low and increase until you reach this level. The other variable is the size of the nozzle. Example you could use a 70nozzle for macrophages and a smaller sized one for T-cells. What type of cells are you using?

[@cacevedo07]

@@themrvims Hi Vimal, thank you for your prompt reply. I'm using HL60. I usually start with a flow rate of 10 and then increase it to 40 or even 50 events. We have just one nozzle in the lab. I bring the cells at 250,000 cells/ml in a 500 uL volume.

[@DavidRondinone]

How was the banana?

[@athind96]

"Eat a banana"

[@joymwangangye4]

Eat a banana lol 😂