tecan GENios, Руководство по эксплуатации

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7. Quality Control
7.3.2 Cross-Talk Test
Cross talk is a measure for increase of blank values ( blank contains pure buffer
and substrate) due to optical leakage from neighboring wells.
•
Greiner 96 well plate, flat bottom, white using black cover plate.
•
A single well (for example: E5) is supplied with 20µl of 0.3 10
-3
activity
units/µl alkaline phosphatase dilution and with 100µl of substrate
•
Fill all other wells with pure buffer and substrate.
1 2 3 4 5 6 7 8 9
10 11 12
A
B
C
D
E
F
G
H
N N
N
N
B B
B
B B B
B
B
B
B
B
B
B
B
B
B
B
B
B
B B
B
B
B
B
B
B
B
B
B
B B
B B
B
B
B B
B
B
B
B
B
B
B
B B
B
B
B
B
B
B
B
B
B
B
B
B
B
B
B
B B
B
B
B
B
B
B
B
B
B
B
B
B
B
B
B
B
B B
B
B
B
B
B
B
B
B
B
•
Select maximum dynamic range and adjust gain to get a
RLU (Relative Luminescence Units) reading greater than 10
6
.
Measurement should be performed 10 minutes after preparation but not later
than 30 minutes.
Calculation method :
Compute MVB ( B ) from all wells except the nearest neighbor wells ( N ) and the
well filled with alkaline phosphatase (for example: E5).
Calculate the SD (Standard Deviation) of MVB. 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 ) / RLU reading
CT is expected to be
≤
0.0001
June 2002 Operating Manual for GENios, GENios FL, and GENios Plus No: I 112 904 Rev No: 1.1 7-5