Fluorolog-3
This manual is meant to complement your training and is not intended to replace on-the-job training.
WPC Activities Associated
MF-0006; Lasers for Fluorolog (only required for laser use)
Training Contacts
Tracy Mattox (fluorescence, lifetime, quantum yield, 3D)
tmmattox@lbl.gov
67-4111
x2649
Emory Chan (MicroMax plate reader, laser use)
emchan@lbl.gov
67-4114
x7373
Last Updated
10/12/2016
OPERATING PROCEDURE
FLUORESCENCE IN UV/VIS AND IR REGIONS
UV-Vis (CCD) Range: 290nm – 850nm (exit = Axial)
UV/Vis (PMT) Range: 290nm – 850nm (exit = Lateral)
IR Detector Range: 900-2000nm
(850-1600nm available, see Tracy for details)
Lamp/Grating Excitation: 200nm – 640nm
1) **SIGN THE LOG BOOK**
2) Turn on both computers (one under Fluorolog and one on desk) if not already on and wait until they completely boot before proceeding.
3) If using the Xenon lamp, flip the main switch on followed by the lamp. Wait a few minutes for the lamp to warm up before collecting data.
4) Turn on the CCD power supply and Fluorohub
5) Start the Fluorescence software (located on the desktop)
6) Click on the method button to initialize the system. Once initialized, this button reloads the default parameters.
7) Select “Spectra” from the experiment menu:
8) Select the experiment type desired (emission or excitation):
9) The experiment window will appear. Load an established method or adjust the parameters as desired.
10) Instructions on how to set up the software to collect data on all three detectors is listed below. All descriptions are for Emission scans.
Note that Excitation scans are set up in the same manner, but a range of wavelengths is entered into the Excitation window (Excitation 1) and one fixed wavelength is set for the Emission (Triax320 or Emission 2).
OPTION 1 = UV-Vis using PMT Detector
Select Detectors button on left side of screen
Data Storage Window:
Enter name of sample – this will be the file name, and advances by one whole number as data collects
Signals Window:
Enter Integration Time (must be > 0.01s)
Enable the S2 detector (units CPS = counts per second) with a checkmark
Be sure that only one detector is selected
Select Monos button on left side of screen
Emission 2 Window:
Deactivate the IR detector (select “Park” and deselect “Activate”)
Triax320 Window:
Select “Set as Reference”
Set Exit to “Lateral”
Input desired wavelength range (within range of 290-850nm)
Set “Inc” (increments or step, i.e. 1 = collect data every 1nm)
Set “Slit” (slit width)
Excitation 1 Window:
Enter “Wavelength” you wish to excite at (xenon lamp with available gratings allows 200-640nm)
Set “Slit” (slit width)
OPTION 2 = UV-Vis using CCD Detector
COOL CCD DETECTOR WITH LIQUID NITROGEN for 20min before collecting data. Continue to add liquid nitrogen every 2hrs until you turn off the detector.
Select Detectors button on left side of screen
Data Storage Window:
Enter name of sample – this will be the file name, and advances by one whole number as data collects
Signals Window:
Enter Integration Time (must be > 0.1s)
Enable the CCD detector (units CPS = counts per second) with a checkmark
Be sure that only one detector is selected
Select Monos button on left side of screen
Emission 2 Window:
Deactivate the IR detector (select “Park” and deselect “Activate”)
Triax320 Window:
Select “Set as Reference”
Set Exit to “Axial”
Input desired wavelength range (within range of 290-850nm)
Set “Inc” (increments or step, i.e. 1 = collect data every 1nm)
Set “Slit” (slit width)
Excitation 1 Window:
Enter excitation “Wavelength” (xenon lamp allows 200-640nm)
Set “Slit” (slit width)
OPTION 3 = IR Detector
COOL CCD DETECTOR WITH LIQUID NITROGEN for 20min before powering the detector on by flipping the toggle switch to “on.” Continue to add liquid nitrogen every 2hrs until you turn off the detector.
Select Detectors button on left side of screen
Data Storage Window:
Enter name of sample – this will be the file name, and advances by one whole number as data collects
Signals Window:
Enter Integration Time (must be > 0.1s)
Enable the T1 detector and switch units to V
Be sure that only one detector is selected
Select Monos button on left side of screen
Triax320 Window:
Deactivate the UV-Vis detectors (select “Park” and deselect “Activate”)
Emission 2 Window:
Select “Set as Reference”
Input desired wavelength range (within range of 1600-850nm)
Set “Inc” (increments or step, i.e. 1 = collect data every 1nm)
Set “Slit” (slit width)
Excitation1 Window:
Enter excitation “Wavelength” (xenon lamp allows 200-640nm)
Set “Slit” (slit width)
TURN OFF DETECTOR WHEN YOU ARE FINISHED USING THE INSTRUMENT!
11) When all parameters for the designated detector have been selected, press “Run” on the lower right to start collecting data.
12) Once the scan has been completed you will be prompted to save your data. Click “Browse” and save data to your file folder, located under C:\Documents and Settings\Default User.DEFAULT-96070DE\My Documents\Fluorolog data
13) In order to zoom in or determine points on the graph you must first double click on the image.
14) The tabs at the bottom of the page may be selected to view numerical data and any mapping information.
15) Helpful Keys:
NOTE: In order to manipulate the graph - zoom in, get accurate x,y coordinates, you must double left click on the graph!
16) If you are finished with the xenon lamp turn off the lamp and WAIT 10 MINUTES before turning off the Main switch.
17) Turn off the Fluorohub and CCD power supply when finished (both computers should remain on).
18) Write any comments in the log book and notify Tracy Mattox (tmmattox@lbl.gov) immediately if you encounter any problems with the instrument.