tecan infinite 200 - Page 131

8. Quality Control
2008-07 Instructions for Use for infinite® 200 No. 30017581 Rev. No. 1.4 131
Uniformity
Perform the following measurement to determine the Uniformity:
Measurement Parameters:
Parameter Setting
Reading Mode Fluorescence Top
Exc Wavelength
infinite
®
F200: 485 (20 nm)
infinite
®
M200: 485 nm
Em Wavelength
infinite
®
F200: 535 (25 nm)
infinite
®
M200: 535 nm
Number of flashes 25
Integration Time 40
Settle Time 0
Gain Optimal
Plate Type GRE96fb
Plate Layout:
Pipette 200 µl of 1 nm Fluorescein or the blank solution (0.01 M NaOH) into the
appropriate wells according to the plate layout:
<> 1 2 3 4 5 6 7 8 9 10 11 12
A
B
C
D
E
F
G
H
1 nm Fluorescein
Blank
1 nm Fluorescein
Blank
1 nm Fluorescein
Blank
1 nm Fluorescein
Blank
1 nm Fluorescein
Blank
1 nm Fluorescein
Blank
Material/Reagents:
1 nm Fluorescein (in 0.01 M NaOH) (Fluorescein sodium salt, Sigma F6377)
0.01 M NaOH (=Blank) (NaOH pellets, Merck article no. 6495 or Sigma S8045)
1 Greiner 96-well plate black
200 µl Pipette + tips
Calculation of Uniformity:
F
F
mean
100*stdev
(%)Uniformity =
mean
F
Average RFU value of wells filled with fluorophore
stdev
F
Standard deviation of RFU values of wells filled with fluorophore
The result of the formula determines the uniformity in % CV.