Biotek Synergy H1, Руководство оператора

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Results Analysis
The Absorbance Liquid Test procedures begin on page 114 .
Absorbance specifications used with the liquid tests:
Accuracy:
± 1.0% ± 0.010 OD from 0.000 to 2.000 OD
± 3.0% ± 0.010 OD from 2.000 OD to 2.500 OD
Repeatability:
± 1.0% ± 0.005 OD from 0.000 to 2.000 OD
± 3.0% ± 0.005 OD from 2.000 OD to 2.500 OD
Absorbance Liquid Test 1
1. The plate is read five times in the “Normal” position at 405 nm. Calculate the Mean
OD and Standard Deviation of those five reads for each well in columns 1 and 2.
2. For each well in columns 1 and 2, calculate the Allowed Deviation using the
Repeatability specification for a 96-well plate (Mean OD x 0.010 + 0.005). For
each well, its Standard Deviation should be less than its Allowed Deviation.
Example: Five readings in well A1 of 0.802, 0.802, 0.799, 0.798, and 0.801 result
in a Mean of 0.8004 and a Standard Deviation of 0.0018. The Mean multiplied by
1.0% (0.8004 * 0.010) equals 0.008, and when added to 0.005 equals 0.013; this is
the Allowed Deviation for well A1. Since the Standard Deviation for well A1 is less
than 0.013, the well meets the test criteria.
3. The plate is read five times in the “Turnaround” position at 405 nm. Calculate the
Mean OD of those five reads for each well in columns 11 and 12.
4. Perform a mathematical comparison of the Mean values for each well in its
Normal and Turnaround positions (that is, compare A1 to H12, A2 to H11, B1 to
G12,… H2 to A11). To pass the test, the differences in the compared Mean values
must be within the Accuracy specification for a 96-well microplate.
Example: If the Mean value for well A1 in the Normal position is 1.902 with a
specified accuracy of ±1.0% ±0.010 OD, then the expected range for the Mean of
the well in its Turnaround (H12) position is 1.873 to 1.931 OD. 1.902 x 0.010 +
0.010 = 0.029; 1.902 - 0.029 = 1.873; 1.902 + 0.029 = 1.931.
Absorbance Liquid Test 2
1. The plate is read five times at 450/630 nm ("Normal" position), resulting in five
sets of Delta OD data. Calculate results for Linearity:
o
Calculate the mean absorbance for each well, and average the means for each
concentration.
o
Perform a regression analysis on the data to determine if there is adequate
linearity. Since it is somewhat difficult to achieve high pipetting accuracy when
80 | Chapter 7: Instrument Qualification Process
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