Re: FMO - compensation

You cannot use FMO's to compensate.  Compensation is an 
instrumentation issue and requires single-stained samples to 
calibrate the spillover.  FMO's are used to identify how to analyze 
the sample, i.e., to assist with gate setting.

Your boss is not understanding compensation; please direct him to 
resources like my web site (<http://www.drmr.com/compensation), or 
various articles in Cytometry, Current Protocols, etc.

mr



At 3:49 PM -0500 12/2/04, Lowe, Kimberley wrote:
>Hi there,
>Just a quick question. Our group has been doing 'compensation' with FMOs
>(Flourescence Minus One).  I have been saying that we really need single
>positive controls to do the compensation accurately, but, because we 
>are looking
>for very rare cells, my boss has argued that they want to take into 
>account all
>the color interactions when compensating.  It is very difficult (for me) to do
>compensation this way when I have been use to using single colors. What do
>others think?	We are looking for a population that is .01 % of the entire
>population and we are using 4 colors - Fitc, PE, PerCP and APC on a two laser
>Calibur.
>Thanks so much for your responses!
>Cheers,
>Kim Lowe
>Childrens Hospital
>Boston, MA