tecan Infinite 200 PRO - Calculation of Fluorescence Polarization Parameters

5. Operating the Instrument
2016-04 IFU for Infinite 200 PRO No. 30052730 Rev. No. 1.6 101
5.6.8 Calculation of Fluorescence Polarization Parameters
G-Factor:
))(1(
))(1(
par
buf
par
ref
ref
cross
buf
cross
ref
ref
RFURFUP
RFURFUP
G
−−
−+
=
[ ]
bufferofunitscefluorescenrelativeAveragedRFU
referenceofunitscefluorescenrelativeAveragedRFU
referenceofvalueonPolarizatiP
buf
ref
ref
...
...
P ...
Blank Reduction:
The mean value of the respective blank wells is subtracted from each value.
welleachfor
RFURFU
RFURFU
RFURFU
RFURFU
RFU
par
blk
par
blk
par
blk
par
smp
par
buf
par
buf
par
buf
par
ref
par







−
−
−
−
=∆
welleachfor
RFURFU
RFURFU
RFURFU
RFURFU
RFU
cross
blk
cross
blk
cross
blk
cross
smp
cross
buf
cross
buf
cross
buf
cross
ref
cross







−
−
−
−
=∆
Intensities:
Parallel and perpendicular intensities are calculated using the following formulas:
parpar
RFUGI ∆= *
crosscross
RFUI ∆=
Polarization:
crosspar
crosspar
II
II
P
+
−
=
Anisotropy:
crosspar
crosspar
II
II
A
*2+
−
=
Total Intensity:
crosspar
tot
III *2+=