Perkin-Elmer LS-55 Fluorimeter#
Procedure#
Make sure the instrument is turned on. The switch is in the back.
Log in with password Springchem2020 if needed.
Open BLDevelopment
Go to .
Use the FL Scan Aquisition Module. Everything else should be set to None.
Click the Manual Control Tab and make sure that Comport Com1 is checked. Everything else should be unchecked.
Open the sample compartment by pulling on the top of the light blue box. Notice that a nifty sample changer is installed! Make sure that the fluorescein sample is in the path of the light beam. If not, turn the knob to rotate the samples.
Note
The fourth slot is a little broken, but can be used if you use use the microspatula located on top of the PE LS 55 to pull back the clamp.
Back on the Acquisition tab, make sure that the Scan Type is Emission, the Ex. and Em. slit widths are set to 6 nm, the scan speed is 500 nm/min, and the Gain is set to Low. Click the play button and then Start. Click Stop when the window pops up after your scan.
Tip
If the data doesn’t look great, try making the scan speed slower.
Find the maximum of the peak. This is your Emission Maximum.
Change the Scan Type to Excitation and the Em. WL to the value you determined in the last step. You may need to decrease your slit widths to 2.5. Identify the maximum here. This is your Excitation Maximum.
Finally, change the scan type back to Emission. This time, change your Ex. WL to the value you found in your previous step.
When you are finished observing your spectra, close the program.
If you are the last person to use the instrument, turn it off.
Attention
It is very important to turn this instrument off to extend the life of the bulb.
Questions#
Why did I make you go back and forth between emission and excitation scans?
Is there anything interesting about the emission and excitation spectra? As in, how do the shapes of these spectra appear?
What does slit width do to the data? Why did you need to decrease it?