Standard Operating Procedure -- Operation of a Perkin-Elmer LS 55 Luminescence Spectrometer
1. Turn on the spectrometer and allow it to warm up for ten minutes. Then turn on the computer.
The instrument is on/off switch is located on the left-hand side panel of the instrument. Turning the instrument on the neon lamp located on the front lower left-hand side of the instrument is lit. The LS 55 takes about 1 minute to initalize and should be turn on before the computer. When Windows has loaded start the FL WinLab software.
The menu bar allows the following selections. The applications tool bar contains icons as shortcuts for applications. At the bottom of the screen is a the menu showing the current Status Line.
2. Open FL WinLab software from desktop icon.
3. Check the status of the instrument by clicking Application, then Status. This page displays a schematic of the instrumental optical system with icons for each individual component. The current status of the instrument is described in text next to the icons. In the Expert Mode the setup can be modified to change between the different modes.
The optical system of the instrument is displayed in a graphical form in the Setup application. By simply clicking on icons, parameters can be easily and directly changed. Parameters can be stored in a method file for later use. In the Intracellular Calibration application, data can be transferred automatically from the sample and calibration runs. The graphical approach of the ICBC module provides you point and click selection of the data range and automatically displays the averaged calibration value of the chosen data.
4. The source light will be on. Do not change any Setup mode/Source parameters for the source. The excitation/emission monochromators can be changed in the Methods section. The only field that may be changed is the emission filter option. Leave ALL Detector parameters unchanged.
5. For Excitation/Emission Scans
Select Application, then Scan from the toolbar. You will see four tabs: Setup Parameters, Realtime Options, User Info, and View Results.
Setup Parameters Tab – For an excitation scan, select the emission wavelength and the range of excitation wavelengths to be scanned. For an emission scan, select the excitation wavelength and the range of emission wavelengths to be scanned. Change excitation and emission slit widths and scan speed if necessary.
6. Realtime Options Tab – Here you can select to automatically subtract a solvent background. Leave response time set to “Auto.”
User Info Tab – Here you can “lock” a method so that it cannot be altered as well as define the scale of the y-axis.
7. View Results Tab – The viewing window for chromatograms is located here.
8. Setup the parameters for your scan and hit the green GO button (from the traffic light signal).
9. For Viewing and Manipulating Spectra.
Chose View from the main toolbar, then New Graph Window. To retrieve a spectrum from the hard drive select File, then open the data file from the appropriate directory.
Use the View menu of the toolbar for manipulations such as adding additional graphs and text.
10. To create printable result graphs, select Data Handling from the toolbar, then Report Builder.
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