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Save the normalization protocol and then check the plate map. Two plates are required for FlashPlate normalization. The first one is an empty plate that is used for determining the background of each of the detectors. For example, a six-detector instrument will store six background values and subtract the appropriate value from each unknown - detector specific background subtraction. The second plate has the active sample in G11 (96-well plate) or M22 (384-well plate). It is advisable to acquire at least 10,000 counts for each reading.
Next, create a counting protocol. The key selections, Other, upper PM, and window setting must be identical to the normalization or the normalization will not be visible from within the counting protocol.
Now, when unknowns are selected, the CPM1 is the raw value and the CCPM1 is corrected for background and efficiency.
To avoid phosphorescence, both normalization plates and assay plates should be dark-adapted for a couple of minutes before counting.
FlashPlate is a registered trademark of Packard Instrument Company, exclusively licensed to PerkinElmer Life Sciences, Inc. FlashPlate is protected under US Patent 5,496,502 and foreign equivalents, to all of which PerkinElmer Life Sciences holds an exclusive worldwide license. TopCount is a registered trademark of Packard Instrument Co.