Precision Test; Cross-Talk Test - tecan GENios Operating Manual

7.2.3

Precision Test

7.2.4

Cross-Talk Test

June 2002 Operating Manual for GENios, GENios FL, and GENios Plus No: I 112 904 Rev No: 1.1
•
Use the same plate as mentioned in 7.2.2.
•
Perform 7 measurements with number of flashes set to 10, gain as above.
Compute the CV (Coefficient of Variation) value of each well in column 4.
The CV value should be <=2.5%.
In fluorescence mode, excitation and emission is performed in the center of each
well. The cross-talk test helps to identify when the excitation light hits a
neighboring well and leads to a false fluorescence intensity.
•
Use a 96 well plate, for example: a Greiner flat bottom, black microplate.
Volume per well - 200 µl
•
Isolated wells E2, E5, E8, E11 contain 10 µM Fluorescein
•
Prepare a 1:1 dilution series in row A from A1 to A12 starting with 1µM
Fluorescein
•
Fill the wells with buffer that are indicated with the letters N and B.
4 5
1 2 3
A
B
B
B B B B
C
N
N
D
N N
N
E
N N
F
B
B B B B
G
B B B B B
H
•
Select extended dynamic range and perform a measurement with 10 flashes
Compute the MVB (Mean Value of Blanks), using the wells indicated with the
letter B and the SD (Standard Deviation). Use factor 3 to get higher statistical
reliability.
Calculate the MVN (Mean Value of the Neighbors), using the wells indicated with
the letter N.
If MVN is within the interval MVB +/- 3 SD, cross talk cannot be clearly
distinguished from the background.
If MVN is higher than MVB +3 SD the cross talk CT (Cross Talk) calculates
according to:
CT = (MVN – MVB ) / Average RFU (E2, E5, E8, E11).
CT is expected to be ≤ 0.0001.
6 7 8 9
10 11
B
B B B B B
N N
N N N
N
N N
B B B B
B B
B
B B B B
B
7. Quality Control
12
B
N
B
B
7-3