I just discover this série. I really like how clear and concise is the video on one flowjo function. And indicating the other videos on more on the topic
Hello, compliments of the new year to you! Thank you for the video. Is it possible to have the sample ID for each population once concatenated and viewing as - for example - FSC-H on y-axis and Sample ID on x-axis? Also, if you're familiar with the CBA plugin, are you able to please film a video on how to use the plugin and troubleshoot when there's errors? Thank you!
High background refers to the spread in the signal in the unstained sample. The last sample on the plot is unstained and you can see how far the signal spreads up the y axis.
Awesome video! it really explains an easy way to understand how across titrations the different variations of fluorcrome intensity compared to the NS. Could you post a video using the FlowJo Stainindex plug in? I really want it to work and dont know how it really works. Very best
They will be ordered based on the order in which you selected them. You could also give each file a keyword (numerical keyword is recommended) to make it easier in the future
Can you present ab titration protocol : do you start with the manufacturer concentration titer ?and how many dilutions you do and what is the parameter you use (signal/ noise) to chose the optimal ab concentrations.Thank you
Awesome! I'm new in flow cytometry, lots of things to learn. As you said, like the CD11c, we can not see the break in flowjo, have to calculate the SI to find the best concentration to use, could you please tell me how to calculate the SI and make the curve to see which concentration that I can use? Thank you so much.
Hi @chris, I don't normally but I do try run my titrations on populations that I know will have good viability. Then I don't need to worry about compensation. For stain index calculations, I recommend taking a look at this : fluorofinder.com/newsletter-staining-index/
Very useful video! Thanks! I have a question: When I selected Sample ID, nearly all my events were on the chart edges. I had to change into log axis in order to see the events, but the resolution is bad. Is it because how I acquired my samples? Thanks in advance!
I made a mistake in naming my samples while running flow cytometry. I have already analyzed the data using Flow Jo, and are in wsp format. Is there any way to rename the sample names in analyzed data in Flow jo?
The filename from acquisition is hard-written into the FCS file and can't be changed in FlowJo. Your best bet is to use keywords to rename and go based off that. OR you can go back into your acquisition file and rename there and export again. Hope that helps!
Thanks for your comment! This article will give you the formula for the calculation: fluorofinder.com/newsletter-staining-index/ FlowJo also recently released a stain index calculation plug-in… I’ve not yet used it but it seems to be pretty good.
![FlowJo [BASICS]](http://i.ytimg.com/vi/xAWUVy8r9gE/mqdefault.jpg)
![FlowJo [BASICS]](/img/tr.png)
![Adjusting compensation: how to handle a "bad" matrix [FlowJo advanced]](http://i.ytimg.com/vi/NEtDeck_V3I/mqdefault.jpg)
I just discover this série. I really like how clear and concise is the video on one flowjo function. And indicating the other videos on more on the topic
Please could you provide a video in how to calculate the stain index? This video was fantastic, as I literally did a Ab titration last week.
Stain index video filming next week :)
Hello, compliments of the new year to you! Thank you for the video. Is it possible to have the sample ID for each population once concatenated and viewing as - for example - FSC-H on y-axis and Sample ID on x-axis?
Also, if you're familiar with the CBA plugin, are you able to please film a video on how to use the plugin and troubleshoot when there's errors?
Thank you!
@@saieshnipillay9406 yes! You can put the sample ID on whichever axis you prefer.
Unfortunately I have no experience with the CBA plugin
At 2:53, you mentioned that "this particular marker has a high background", how do you mean by that? How do you see the "background"??
High background refers to the spread in the signal in the unstained sample. The last sample on the plot is unstained and you can see how far the signal spreads up the y axis.
Awesome video! it really explains an easy way to understand how across titrations the different variations of fluorcrome intensity compared to the NS. Could you post a video using the FlowJo Stainindex plug in? I really want it to work and dont know how it really works. Very best
Sorry about the delay- I will be filming this next week!
At 2:44 and 3:59 to show concat titration plot, how do you which plot for which titration?
They will be ordered based on the order in which you selected them. You could also give each file a keyword (numerical keyword is recommended) to make it easier in the future
@@ajarieger_flow Thanks for your kind explanation.
Can you present ab titration protocol :
do you start with the manufacturer concentration titer ?and how many dilutions you do and what is the parameter you use (signal/ noise) to chose the optimal ab concentrations.Thank you
That would be a great video- I will add it to my list :)
What if I don't have unstained cells .. no negative control
@@kaybash21 you can treat it the same way, you just won’t have an unstained on your plot. You can still do stain index etc as normal
Awesome! I'm new in flow cytometry, lots of things to learn. As you said, like the CD11c, we can not see the break in flowjo, have to calculate the SI to find the best concentration to use, could you please tell me how to calculate the SI and make the curve to see which concentration that I can use? Thank you so much.
Thanks for tuning in! We just posted a video yesterday on stain index- you can check it out here:
ruclips.net/video/R-BfoEoDNXQ/видео.html
I am working on a follow-up video to show stain index calculation with more challenging populations.
Should you not include a live/Dead with your dilutions, as dead cells stain everything?
Hi @chris, I don't normally but I do try run my titrations on populations that I know will have good viability. Then I don't need to worry about compensation.
For stain index calculations, I recommend taking a look at this : fluorofinder.com/newsletter-staining-index/
I just discovered your channel and I am very glad I did. Really appreciate the effort you put in your videos. Thanks a lot! keep going
Thanks!! If there are any topics you want covered, let me know 😁
Very useful video! Thanks! I have a question: When I selected Sample ID, nearly all my events were on the chart edges. I had to change into log axis in order to see the events, but the resolution is bad. Is it because how I acquired my samples? Thanks in advance!
Thanks! Without seeing the data, it is hard to tell… feel free to send me an email @aja@ualberta.ca and I can take a quick look
@@ajarieger_flow thank you!! Happy Thanksgiving!
I made a mistake in naming my samples while running flow cytometry. I have already analyzed the data using Flow Jo, and are in wsp format. Is there any way to rename the sample names in analyzed data in Flow jo?
The filename from acquisition is hard-written into the FCS file and can't be changed in FlowJo. Your best bet is to use keywords to rename and go based off that. OR you can go back into your acquisition file and rename there and export again. Hope that helps!
Hi Aja. Could you make a new video for flowjo to introduce a statistic analysis if you are free?
Thanks,
Fei
For sure! So I am on the same page- are you looking for a video that shows how to get statistics like gate % and MFI from FlowJo?
@@ajarieger_flow yes, if you are available to do those. Thanks
Very nice video, would be great if you could show us how to calculate the s.i. :)
Thanks for your comment! This article will give you the formula for the calculation: fluorofinder.com/newsletter-staining-index/
FlowJo also recently released a stain index calculation plug-in… I’ve not yet used it but it seems to be pretty good.
I will be filming a stain index video next week :)
Very helpful