Bio-Rad Bio-Plex Software® Upgrades and Conversions - Manual (Page 237)

Normalization Formulas
225
Normalized Fluorescence Values (Norm FI)
(This calculation requires internal controls to be chosen.)
Normalized Fluorescence Values (Norm FI) are fluorescence values adjusted
to account for experimental sample to sample variations (loading differences).
Norm FI can be used to compare results across experiments to get a clearer
picture of experimental variability. It is left to the researcher to decide the
appropriateness of this calculation for a particular experiment.
Norm FI is calculated as follows when housekeeping genes (internal controls)
are chosen.
Norm FI
(well)
= FI Gene of Interest *
where Avg All HKG FI
(ALL wells)
= the average FI for all housekeeping genes in
every well of the experiment.
This number serves as a mean factor around which all others will fall and
converts a “unitless” ratio back into FI.
The next calculation averages per well data to arrive at the per sample
normalized fluorescence value (Norm FI
(sample)
).
Norm FI
(sample)
=
When control is chosen, Norm FI is calculated as follows.
Norm FI
(sample x)
= Norm ratio
(sample x)
* Average FI
(control sample)
Average FI
(control sample)
serves as a mean factor around which all others will
fall and converts a “unitless” ratio back into FI.
BioPlex_6.book Page 225 Friday, September 23, 2011 11:55 AM