Very usefull video. Would´ve been nice to watch the whole procedure, from FSC/SSC --> FL2A/FL2W(or other doublet discrimination method) --> Cell cycle analysis
Thank you, FlowJo. As a further clarification, I think gating the phases for the control for use with other samples should only be applicable if the cells were synchronized when the treatments were carried out. Could you kindly confirm?
Do all samples have to follow the same constaint n value that was done with a negative control or are we allowed to tweak n values per sample to minimize RMSDs?
what happens if the position of G1 shifts between samples and I applied the same model to every sample (restricting G1): For instance, the peak appears in 200 in one sample and in 230 in another sample. I should manually reset the position of said sample, shouldnt I?
Hi user, as long as the peak is still within the G1 gate, the model should work as expected. If the peak has shifted outside of the gate then the G1 gate will need to be shifted to include the G1 peak. If you have more questions please email us at FlowJo@bd.com!
@@FlowJoMediaHi, the suggestion of shifting the gate is contrary to what Jack indicated in the video. The essence of gating is to be able to use the control as a standard to compare with the other tests.
What happens if I try to open up a cell cycle model from my control and it says "could not calculate model: unable to calculate model based on inputs"?
Hi sir: At minute 21:20, you made a statement which is contrary to the data. More cells in G1 phase for the control, but it's not so. How will your clarify this, please?
Hi, For the data Jack presented the cells were released from an inhibitor keeping them in the G1 phase before the experiment began. Because the data was acquired shortly after the cells were released from the G1 phase, most of the cells for the experimental samples were in S phase. The distribution of cells in other cell cycle experiments may look different depending on the inhibitor used and the time at which the data was acquired.
if you guys are going to use shortcuts, you should write them on the video. This should also just be a written tutorial so people don't need to watch videos to use a product
Very usefull video. Would´ve been nice to watch the whole procedure, from FSC/SSC --> FL2A/FL2W(or other doublet discrimination method) --> Cell cycle analysis
Very helpful! Please do more like this! Thanks
Thank you for your video! It was really helpful!
super helpful, thank you so much!
Awesome thanks!
Thank you, FlowJo. As a further clarification, I think gating the phases for the control for use with other samples should only be applicable if the cells were synchronized when the treatments were carried out. Could you kindly confirm?
What if your data doesn’t fit the cell cycle model?
Do all samples have to follow the same constaint n value that was done with a negative control or are we allowed to tweak n values per sample to minimize RMSDs?
is it the same if you stain your cells with propidiumiodid? or is there a different range on the x-axis for the phases?
If i want to change scale bar on x-axis how can i change
what happens if the position of G1 shifts between samples and I applied the same model to every sample (restricting G1): For instance, the peak appears in 200 in one sample and in 230 in another sample. I should manually reset the position of said sample, shouldnt I?
Hi user, as long as the peak is still within the G1 gate, the model should work as expected. If the peak has shifted outside of the gate then the G1 gate will need to be shifted to include the G1 peak. If you have more questions please email us at FlowJo@bd.com!
@@FlowJoMediaHi, the suggestion of shifting the gate is contrary to what Jack indicated in the video. The essence of gating is to be able to use the control as a standard to compare with the other tests.
What may happening in my cell cycle analyses when my cells shows more G2/m than G1? Like it's inversed...
Hi Felipe, please reach out to us at flowjo@bd.com and we can assist you further and discuss some possibilities in this case. Thank you!
What happens if I try to open up a cell cycle model from my control and it says "could not calculate model: unable to calculate model based on inputs"?
do you find out the solution for this case?
Hey, greetings! Did you solve this?
I got it! You have to change the filter to "FSC-A :: FSC-A"
@@vivi7279 Hi! Sorry could you please clarify how you solved the above problem?
Hi sir: At minute 21:20, you made a statement which is contrary to the data. More cells in G1 phase for the control, but it's not so. How will your clarify this, please?
Hi,
For the data Jack presented the cells were released from an inhibitor keeping them in the G1 phase before the experiment began. Because the data was acquired shortly after the cells were released from the G1 phase, most of the cells for the experimental samples were in S phase. The distribution of cells in other cell cycle experiments may look different depending on the inhibitor used and the time at which the data was acquired.
@@FlowJoMedia Many thanks. In addition, it seems single cell discrimination was not carried out after gating out cells from debris.
thanks
if you guys are going to use shortcuts, you should write them on the video. This should also just be a written tutorial so people don't need to watch videos to use a product