Specifications

PN 718113AF
25
Laser Induced Fluorescence Detection
Operation
• Fluorescence yield of the molecule being measured
• Background buffer composition and properties
• Separation parameters (temperature, voltage, etc.)
• Laser power
To minimize variation caused by (small) changes to any of the parameters listed above, a correction
factor can be used to normalize the detector response against a defined standard.
The Calibration Correction Factor (CCF) is the mathematical relationship between the fluorescence
from a known concentration of the analyte and the associated signal detected by the instrument.
Subsequent measurements are multiplied by the factor to determine the relative fluorescence unit.
For the LIF Detector system, a Calibration Wizard is available to simplify the determination of the
calibration correction factor. During the CCF determination, the Calibration Wizard instructs the
instrument to execute the following:
• Measure the background (with buffer)
• Measure the signal from the “standard”
The Calibration Wizard will then:
• Calculate the change in signal between the standard and the background buffer
• Calculate the correction factor using the following equation:
where:
Target RFU value = Target value for the standard solution used (entered by user into the Wizard).
Measured RFU value = Change in signal between background buffer and standard.
CCF
o
= The current value of the Calibration Correction Factor (CCF).
CCF
Target RFU Value
Measured RFU Value
--------------- ------------------- --------------------
CCF
O
•=