gating on histogram overlap

Nebe-Von-Caron, G g.nebe-von-caron at unipath.com
Wed Oct 11 06:09:35 EDT 2006


If you are doing multicolour you have to remember the problem of
'population widening' by the overlap which could screw your gate. Just
as a quick summary on gating in the context of controls and compensation
you might look at a flowjo presentation to appreciate the gating problem
over several colour staining.
http://www.urmc.rochester.edu/wnyfug/pdf/wilshire-2004.pdf

The other thing vividly stuck to my mind is a lecture from good old Jim
Watson at Brian Shenton's flow meeting - mainly because I could
understand him whilst I struggled with a guy from Newcastle trying to
explain to me why I could not understand a word from the guy from
Glasgow, both in their most lovely accent. Jim showed us a series of
histograms to identify the % positive he had simulated from independent
distributions. You might read his book on data analysis or use our ISAC
journal to brush up on the info. 

http://www3.interscience.wiley.com/cgi-bin/abstract/76502809/

As an isac member you should have access to the full journal content.
However, I actually forgot to log in when trying to access the pdf
article above (wouldn't remember my password anyhow), so it looks that
some older material is accessible for free. I hope this is not a glitch
but journal policy, so please use it, cite it and make others aware of
this excellent resource.



Gerhard


> -----Original Message-----
> From: Stevan Lauriault [mailto:stevan at lauriault.com] 
> Sent: 05 October 2006 22:50
> To: Cytometry Mailing List
> Subject: gating on histogram overlap
> 
> 
> Dear All,
> 
> When analyzing histograms of dimly fluorescent samples, and 
> comparing to their specific isotype controls on overlay, 
> there is a significantly large overlap between the two 
> populations (ex. ~10%).  Is there a universally accepted 
> method to statistically quantify the relative proportions of 
> positive vs. negative populations?
> 
> We are unsure whether to gate the entire histogram, or just 
> the positive region when determining relative fluorescence 
> intensity values.
> 
> Any advice would be appreciated.
> 
> Thank you,
> 
> Stevan Lauriault, B.Sc.
> Research Assistant
> Trauma Group
> Operational Medicine Section
> Defence R&D Canada - Toronto
> Stevan.lauriault at drdc-rddc.gc.ca
> Work:  416-635-2000 ext. 2303
> Cell:  416-949-4066 
> 
> 
> 
> 
> ________________________________________________________________
> Sent via the WebMail system at lauriault.com
> 
> 
> 




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