Amazing material and tips! I was working many years ago in this world of flow cytometry and I came back to this four months ago. Needed a refresh and review concepts and I found you! 🙏Very grateful!!! Looking forward for the advanced level and recomendations by using a high multiparametric panel😊🥰
Aja, I cannot thank you enough for these amazing videos. I have to teach myself flow cytometry since there´s no one in my lab who uses it on the level I need it to function and have been searching for a comprehensive, concise and well made explanation and THIS IS IT. I´m studying these videos like a bible and I´m incredibly thankful. You´re amazing and I am looking forward to any other content!!!
Thank you for your great videos! Even though I've been doing flow cytometry for a long time I still picked up a couple of pointers from your explanations.
I need to setup a 20 color panel for 1 sample tube. Im going to be using Beckman Coulter's Cytoflex LX. Can someone tell me which flourochromes I should not mix together? I know FITC and AF488 should not be mixed because they have similar emission and excitation specs. I also don't like using BV510 because it is so weak and interfere with a few other flourochromes. Can someone help me?
If your facility has the option, look into Spectral Flow Cytometry. It's a lot better at unmixing similar flourochromes than traditional flow cytometry. BV510 is even brighter.
Yes you should still for any viability dye. For these controls you will want to use cells that are somewhat dead... heat shock, repeated freeze/thaw, or addition of chemicals that induce cell death are all common methods here
It will ultimately depend on the optical configuration of your instrument- but if all lasers are spatially separated, it should work. I would however recommend that you don’t match them with markers on the same cell type
Thanks for the informative series, I really appreciate it, but could you pls avoid using the background music in the future as it makes the voice less clear. Thnx again.
![Adjusting compensation: how to handle a "bad" matrix [FlowJo advanced]](/img/n.gif)
Amazing material and tips! I was working many years ago in this world of flow cytometry and I came back to this four months ago. Needed a refresh and review concepts and I found you! 🙏Very grateful!!! Looking forward for the advanced level and recomendations by using a high multiparametric panel😊🥰
The antigen hierarchy is such a elegant and easy to implement tip! Thanks a lot.
Aja, I cannot thank you enough for these amazing videos.
I have to teach myself flow cytometry since there´s no one in my lab who uses it on the level I need it to function and have been searching for a comprehensive, concise and well made explanation and THIS IS IT.
I´m studying these videos like a bible and I´m incredibly thankful.
You´re amazing and I am looking forward to any other content!!!
I’m glad they’ve been helpful!!
Thank you for your great videos! Even though I've been doing flow cytometry for a long time I still picked up a couple of pointers from your explanations.
Glad you found it helpful!
Excellent videos, thanks. Would love to see the advanced panel design video.
Noted! I am planning some new videos to release soon- I will add this to my list
your videos are amazing thanks for the geat quality
There's definitely an art to it. “Sometimes science is more art than science, Morty. Lot of people don't get that.” - Rick Sanchez
Great! Thanks
I need to setup a 20 color panel for 1 sample tube. Im going to be using Beckman Coulter's Cytoflex LX. Can someone tell me which flourochromes I should not mix together? I know FITC and AF488 should not be mixed because they have similar emission and excitation specs. I also don't like using BV510 because it is so weak and interfere with a few other flourochromes. Can someone help me?
If your facility has the option, look into Spectral Flow Cytometry. It's a lot better at unmixing similar flourochromes than traditional flow cytometry. BV510 is even brighter.
Thank you so much!
Do we need to make compensation controls for Dapi? Assuming most cells are alive, how can I have enough events for dapi +ve cells.?
Yes you should still for any viability dye. For these controls you will want to use cells that are somewhat dead... heat shock, repeated freeze/thaw, or addition of chemicals that induce cell death are all common methods here
Can I use BB700 and BV711 in the same panel? The filters are the same (710/50) but in different lasers.
It will ultimately depend on the optical configuration of your instrument- but if all lasers are spatially separated, it should work. I would however recommend that you don’t match them with markers on the same cell type
Thanks for the informative series, I really appreciate it, but could you pls avoid using the background music in the future as it makes the voice less clear.
Thnx again.