Biotek Synergy H1, Operator'S Manual

Page: 124 / 190

Parameter Setting
Gain Auto, Scale to High Wells, C1, 80000
Read Speed Normal
Delay after plate movement 350 msec
Measurements per data point 100
Lamp Energy Low (faster)
Read Height 7.00 mm
Results Analysis
The Fluorescence Liquid Test procedures begin on page 121 .
Corners Test
1. Calculate the Mean of the 12 "corner" wells (A1–A3, A10–A12, H1–H3, and H10–
H12).
2. Calculate the Standard Deviation of the same 12 wells.
3. Calculate the %CV: (Standard Deviation/Mean)*100
The %CV must be <3.0 to pass.
Sensitivity Test
1. Calculate the Mean and Standard Deviation of the 16 reads for each of the buffer
wells (C9, D9, E9).
2. Among the three buffer wells, find the Median Standard Deviation and
corresponding Mean.
3. Calculate the Mean for the 16 reads of the SF (or MUB) Concentration well (D7).
4. Calculate the Signal-to-Noise Ratio (SNR) using the Mean SF (or MUB)
Concentration, Buffer Median STD with its corresponding Buffer Mean:
(<SF or MUB>Mean-BufferMean)/(3*BufferSTD)
5. Calculate the Detection Limit:
Sodium Fluorescein: Using the known concentration value of SF and the
calculated SNR: 1000/SNR
o
Filter-based system, top optics: The Detection Limit must be <=10 pM (2
pg/mL) to pass.
o
Monochromator-based system, top/bottom optics: The Detection Limit must be
<=20 pM (3.76 pg/mL) to pass.
Methylumbelliferone: Using the known concentration value of MUB and the
calculated SNR: 17.6/SNR
98 | Chapter 7: Instrument Qualification Process
BioTek Instruments, Inc.