Chem 201 DE Matthews page 1

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Instructions for Using the Photon Technologies Fluorimeter

The following document is a supplemental guide intended to be used by CHEM 201 students for
the emission and excitation spectroscopy portion of the Molecular Spectroscopy lab. The
procedure described below is for using the Photon Technologies Inc. fluorimeter and its PC data
system.

Getting Going:
1. Bring a flash drive to the lab with you. If you don’t have a flash drive, see your TA to
borrow one.
2. Room A123 door code: _____ (get from TA)
3. Before going to the next step make sure the computer to the fluorimeter is off, AND
make sure the two shutters are CLOSED.
4. Press power button on lamp power supply, then hold down Ignite button until the display
reads between 60-80 Watts.
5. Moving from the top to the bottom, switch on Power Driver and the Power on very
bottom of power supply stack.
6. Once the light source is steady (i.e. between 60-80 W and ~11 V without significant
fluctuations)
a. Open shutters (2) by sliding.
b. Make sure the photomultiplier tube (PMT) is on. (Please do not make any
adjustments to the PMT, if the voltage is too low the TA will adjust it.)
7. Turn on the PC computer.
8. PC computer
a. Login: _____ (get from TA)
b. Password: ______ (get from TA)
9. Programs  FelixGX  Administrator
a. Username: pti
b. Password: ______ (get from TA)

Emission (Fluorescence) Spectrum:

1. Felix32GX  Set Up  “Emission Scan”
2. Select “enable” then choose “emCorri” under the real-time corrections tab
a. Make sure that the slit widths in the emission scan setup are set at 1nm. They will be
preset to this value; you just have to type in “1 nm” for each of the slit widths. Do
not adjust the slits.
3. Obtain the background correction:
a. First, close the two slides, and click on the “Close shutters” button in the software.
b. Rinse the cuvette with the solvent (i.e. n-heptane) ~x3.
i. DO NOT USE ACETONE; a possible alternative solvent would be i-propanol if a
cell needs cleaning.
c. Fill the cuvette with the solvent (i.e. n-heptane). Place cuvette in sample
compartment and close lid.
d. There must be at least 25-nm gap between the excitation and the emission
wavelengths or the detector will overload! For the emission spectrum: look at the
absorbance spectrum and find a maximum (e.g. for a strong peak at 355.0 nm a good
setting would be Excitation 1 = 355.0 nm and Emission 1=380.0 to 500.0 nm).
e. Click Acquire – note that the wavelengths are being set
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f. After, make sure that the “Use” box is checked. Close shutters (manually and in the
software) and remove the cuvette containing n-heptane.
4. Obtain the emission (fluorescence) spectrum of the anthracene:
a. Rinse a cuvette with small amounts of anthracene solution ~3 times. Fill the cuvette
with the anthracene solution and place in fluorimeter.
b. Remember, the “use” box should be checked. Re-open the shutters (manually and in
the software).
c. Use the same excitation and emission settings as in the prior step.
d. Click Start
e. Save the spectrum by selecting it from the list on the left-hand side of the window
and copy and paste the data into a Notepad (.txt) file.
5. Save the data onto a flash-drive.

Excitation Spectrum:
1. Repeat Steps 1 & 2 above for emission scan, except choose “Excitation Scan” and use
“excorr” for the real-time correction.
2. Background correction: Repeat step 3 above.
a. Remember the 25-nm gap between excitation and emission! For example, if you
observe a strong peak at 400.0 nm in the emission spectrum of anthracene, you could
set emission to 400.0 nm and scan excitation from 200-375 nm, but NO higher.
3. Obtain the excitation spectrum of the anthracene as per step 4 for the emission spectrum.
4. To save the data follow steps as shown above.

Shutting Down
1. Turn off the computer & monitor first (make sure it has completely powered down)
2. Turn off the PMT via bottom box in tower
3. Close the slits
4. Turn off the lamp power supply, then switch off the Power on very bottom of power
supply stack, and switch off the Power Driver
5. Replace the plastic covering over all of the instruments (including the computer) to
prevent dust and other particulates from damaging the machines.