[Cytometry] Summary: Indo-1 kinetics, DIVA and Flowjo
simm at treestar.com
Thu Sep 30 11:57:16 EDT 2010
this is a known issue discussed below with a workaround.
FlowJo Support & Development
On Sep 30, 2010, at 1:08 AM, Rosemary Clarke wrote:
> Thanks to everyone who replied to my post:
> Having recently dumped our old LSR1 and acquired a new Fortessa, I am
> currently trying to look at calcium flux using Indo-1 and DIVA for
> the first time. It's the type of analysis I have done hundreds of
> times before and the experiment is working well. The data I get in
> DIVA looks as expected. However, I am having issues when I transfer
> my FCS files (3.0 format) from DIVA into Flowjo. When in DIVA the
> blue/green ratio looks fine (linear) but when the data gets into
> FlowJo it automatically gives it a log scale and it is just wrong and
> can't be analysed. Any clues? Is there something I can do in Flowjo
> or DIVA to stop this from happening?
> As it turns out, simply changing the preferences from displaying log data to linear does not work where ratios are concerned. It seems that ratios produced in DIVA do not transfer to Flowjo. There were a few suggestions to over come this but the one that seemed to work for me was as follows, kindly provided by Cheryl Kim:
> "For some reason the ratio doesn't carryover from Diva so you have to go and create a ratio using the Platform < Derived Parameters function. I've attached a picture of this prompt, most important is the - display axis of lowest and highest value... you can keep playing with these values to make it look more normal. You just click on Add Algebraic Parameter and name it Ratio and this should do
> the trick."
> Thanks again to everyone who responded.
> Dr Rosie Clarke
> Division of Cell Biology and Immunology
> University of Dundee
> Tel: (01382) 384222
> E-mail: r.z.clarke at dundee.ac.uk
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