60
| Instrument Testing
2.
If you have not already done so, create the protocol described starting on
page 61.
3.
Create a new experiment in Gen5 using the
AF_Reliability_<x>.prt
protocol,
with <x> representing the objective you are using for each test.
4.
Click
Plate > Read Plate
, save the experiment, then click
OK
.
5.
When the read is finished, analyze the results as described below.
Analyze the Results
1.
For the set of 100 data points in well F7, calculate the Mean, Slope,
and Intercept. (If using Microsoft Excel, use the SLOPE and INTERCEPT
functions.)
2.
For each data point, calculate the Residual:
[value]–((Slope*[data point sequence #])+Intercept)
Example: 325–((0*1)+325.9)=–0.9 Residual for data point #1
3.
For each data point, compare the absolute value of the Residual with the
objective's Limit (µm) value in the chart below (e.g., 95.4 for 2.5X). If the
absolute value of the Residual is greater than the Limit, note "1" for that data
point; otherwise, note "0".
Depth of Field
Objective
Magnification NA e (µm)
λ (nm)
DOF (µm) Limit (µm)
10X
10
0.3
6.45
377
6.3
12.7
4X
4
0.13
6.45
377
34.7
69.4
Zeiss 2.5X
2.74
0.12
6.45
377
45.8
91.6
Meiji 2.5X
2.25
0.07
6.45
377
117.9
235.8
4.
After conducting this comparison for all 100 data points, add up the
notations and divide the sum by 100; this is the Measured Autofocus Failure
Rate. The Measured Autofocus Failure Rate must be ≤ 1.0% to Pass.
5.
In the Gen5 plate matrix, select the
FM Ratio [0:DAPI 377,447]
data set.
Refer to the chart below for passing criteria for the value displayed in well
F7.
Objective
FM Ratio must be
Meiji 2.5X
> 1.4
Zeiss 2.5X
> 1.4
4X
> 25
10X
> 50
BioTek Instruments, Inc.
Summary of Contents for Cytation 5
Page 13: ...Installation ...
Page 31: ...14 Install the Imager Module 19 Cytation 5 ...
Page 37: ...Getting Started ...
Page 54: ...42 Getting Started BioTek Instruments Inc ...
Page 55: ...Maintenance ...
Page 66: ...54 Maintenance BioTek Instruments Inc ...
Page 67: ...Instrument Testing ...
Page 113: ...Specifications ...
