Thank you for your tuto. I have a question about your FMOs used to determine the positive gates for IFN, Perf etc. Have they been done with isotype antibodies? Thank you
Hi user, No. Those Fluorescent Minus One (FMO) controls do not contain any isotype antibodies. Those FMOs contain all of the stains used in the experiment except for the stain referenced in the FMO, ie the IFN FMO has all of the stains except for the IFN stain. We hope this helps!
Hi Jennifer, Some Cytometers do not display record a time parameter. FlowJo does have a Event Number parameter that can function similar to time, displaying the sequence the events were recorded into the file by the Cytometer. This can be found under the preference “Heart icon” in the “Graphs” section. Hope this helps!
Amazing Tutorial, Thank you so much! I have a question about performance of FlowJo. I got Problems when applying gates to other samples. My FlowJo crashes or goes in a state of infinite loading time. I already set the memory use to 1/4of my RAM. Any suggestions? thanks again! :)
Hi user, The performance may be boosted if the performance preferences are edited. These preferences can be found under the heart shaped icon and then choosing “Performance”. For Windows users the “Max Sample Cache Size” may be lower than optimal. If many gigabytes of data are being loaded into the workspace a higher sample cache size may be more optimal. Here are some suggested sample cache sizes: For 8 GB of RAM ~2000 For 16 GB of RAM ~8000 For 32 GB of RAM ~16000 For 64 GB of RAM ~32000 Issues can occur when the .wsp or .fcs files being used in the workspace are stored on a server. This is because the computer can often have poor or inconsistent connection to files stored in remote locations. If the files are stored on a server, I would try moving them locally (ie to the desktop) and see if that helps. Below is a link to more information on using FlowJo with a remote drive. docs.flowjo.com/flowjo/advanced-features/remote-data/
A whole lot of useful info. Thanks!
Thank you for this! Very helpful!
Thank you so much, very useful.
Glad it was helpful!
Thank you❤❤
This is nice and very informative. is there anyway where i can download flowjo to use for my studies
Thank you for your tuto. I have a question about your FMOs used to determine the positive gates for IFN, Perf etc. Have they been done with isotype antibodies? Thank you
Hi user,
No. Those Fluorescent Minus One (FMO) controls do not contain any isotype antibodies. Those FMOs contain all of the stains used in the experiment except for the stain referenced in the FMO, ie the IFN FMO has all of the stains except for the IFN stain.
We hope this helps!
X axis has no "Time" option for my graphs, is this a preference I can select somewhere? Using Flowjo V10.8
Hi Jennifer,
Some Cytometers do not display record a time parameter. FlowJo does have a Event Number parameter that can function similar to time, displaying the sequence the events were recorded into the file by the Cytometer. This can be found under the preference “Heart icon” in the “Graphs” section. Hope this helps!
Amazing Tutorial, Thank you so much! I have a question about performance of FlowJo. I got Problems when applying gates to other samples. My FlowJo crashes or goes in a state of infinite loading time. I already set the memory use to 1/4of my RAM. Any suggestions? thanks again! :)
Hi user,
The performance may be boosted if the performance preferences are edited. These preferences can be found under the heart shaped icon and then choosing “Performance”.
For Windows users the “Max Sample Cache Size” may be lower than optimal. If many gigabytes of data are being loaded into the workspace a higher sample cache size may be more optimal.
Here are some suggested sample cache sizes:
For 8 GB of RAM ~2000
For 16 GB of RAM ~8000
For 32 GB of RAM ~16000
For 64 GB of RAM ~32000
Issues can occur when the .wsp or .fcs files being used in the workspace are stored on a server. This is because the computer can often have poor or inconsistent connection to files stored in remote locations. If the files are stored on a server, I would try moving them locally (ie to the desktop) and see if that helps. Below is a link to more information on using FlowJo with a remote drive.
docs.flowjo.com/flowjo/advanced-features/remote-data/