Re: MESF and correcting for isotype matched controls

From: Steve Perfetto (sperfetto@pasteur.hjf.org)
Date: Fri Jan 07 2000 - 14:33:57 EST


Calman,

I absolutely agree.  The same type of bias can be introduced in subtracting out
instrument noise since this is essentially instrument specific it would allow
for huge interlaboratory variation.  For this reason we always report the raw
values without subtraction based on the Quantum Simply Cellular system. 
However,  if a low cell control can be standardized to reference the low end as
well as a high end cell control than these values should be valid for
subtraction.  Currently such a control does not exist.

Stephen P. Perfetto, MS.,MT. (ASCP)
Walter Reed Army Institute of Research
Department of Molecular Diagnostics and Pathogenesis
1600 East Gude Drive
Rockville, MD. 20850
_______________________________________________________________________________
Subject: MESF and correcting for isotype matched controls
From:    Calman Prussin <CPRUSSIN@niaid.nih.gov> at Internet_Gateway
Date:    1/6/00  11:42 AM


We are quantitating cell associated antigens using standardized beads and
then generating an MESF from the standard curve generated from the beads.
The question has come up whether or not to "correct" the MESF values
obtained from the specific antibody by subtracting the MESF values obtained
for the  isotype matched controls. 

The problem with correcting the values is that it has little effect on the
samples with high density of Ag, but has a large effect on those with low
expression. As such, if we run replicate samples   on the low expressing
samples we find a larger amount of  variation in the corrected MESFs. My
bias is not to use it, as it seems to introduce more "noise" into the
system. Your thoughts?

Thanks!

> _______________________
> Calman Prussin
> Laboratory of Allergic Diseases
> NIAID/ National Institutes of Health
> 



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