FlowJo on Mac - biexponential adjustment - (Aug/15/2011 )
Hi, I'm using FlowJo 8.7 on Mac to analyze my cells. I'm using a 12 colour stain, and I've used Define Biexponential Transformation in newer editions of FlowJo so that my cells are not sitting on the axis. However, I find that in this version of FlowJo 8.7, all my colours must subscribe to the same settings under biexponential transformation. For example, I cannot use 1 extra negative decade for one colour (eg FITC), and no extra negative decades for another colour (eg PerCP). Is there someway I can get around this? Does it have to do with the number of windows I have open? The number of extra negative decades I show affects how condensed or spread out my cells are and it really helps to be able to choose different settings for different colour channels.
My cells were acquired on BD FACSDiva, and I'm using a width basis of -100. Under Workspace Preferences I have ticked the box to "Allow Custom Visualisation".
If anyone is familiar with FlowJo 8.7 on Mac and knows some way to get around this, please let me know! Thanks much!
I'm sorry because I can only tell you about flowJo 9.2 (I'm not remembering how it was with 8.7). So at least in 9.2, you can manually setup the bi-expo parameters for every channel independently. For that, you have to open one dot plot or histogram and then go plateform / biexponential transformation / manually specify transformation. You will be able to modify the width, the number of positive decades and the number of additional negative decades. If you change the parameters, the modifications will be applied to every samples using the same compensation matrix.
Of note, the biexponantial 'mathematical function' of FlowJo is different than the one used by Diva and is definitely not as good, often leading to weird graphical representations (cells can still look like stuck on a baseline) but still accurate though.
emilytxz on Mon Aug 15 15:37:36 2011 said:
My cells were acquired on BD FACSDiva, and I'm using a width basis of -100. Under Workspace Preferences I have ticked the box to "Allow Custom Visualisation".
Herein lies the problem. FACSDiVa uses a width basis default of -100, while FlowJo uses a default of -10. I do know you can change the width basis in the preferences using the button called 32-bit data (or something like that).